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Integrative Protocol in the Treatment of Rhabdomyosarcoma
Natural Strategies in Collaborative Cancer Care
Compiled by John Connor, M.Ac., L.Ac.
May 23, 2010
Outline
Cells in our body are dynamic. They are very active 24 hours a day, and many of them are constantly dividing. Incorporations of bad mutations during a vulnerable phase in the life of a cell are few, but they do happen. When enough mutations have occurred, the result can be cancer. The modifications of molecules such as DNA, proteins, lipids, etc. by reactive oxygen species (ROS) is the first step in the initiation of several diseases, including cancer as well as cardiovascular, neurodegenerative, and inflammatory diseases. The human body has an immune system that is designed to counteract disease. Unfortunately, conventional cancer therapies compromise the body’s immune system, and this compromised immunity may promote tumorigenesis. (Bagchi and Preuss pp. 343 & 253)
Our objective is to work with your oncologist to formulate a targeted integrative treatment protocol which strives to implement the following:
1. To strengthen the patient with herbs, botanicals and nutrients.
2. To reduce genetic instability by reducing oxidative stress, such as reactive oxygen species.
3. To inhibit abnormal expression of genes by normalizing the function of transcription factors. (For more details see section on Understanding Transcription Factors.)
4. To inhibit abnormal signal transduction, eg. by blocking kinase enzymes and ras signaling. (Martinelli et al 2009)
5. To encourage normal cell to cell communication
6. To inhibit tumor angiogenesis
7. To inhibit invasion
8. To inhibit metastasis
9. To increase the immune response
For example, curcumin has been shown to inhibit the growth of rhabdomyosarcoma cells. The data suggest that curcumin may execute its anticancer activity primarily by blocking mTOR-mediated signaling pathways in the tumor cells. (Beevers et al 2006)
Another study showed that resveratrol exerts a strong inhibition of rhabdomyosarcoma cell proliferation in part by arresting cells in S/G2 phase of the cell cycle. (Chow et al 2005)
It has also been found that resveratrol causes COX-2- and p53-dependent apoptosis (programmed cell death) in head and neck squamous cell cancer cells. (Lin et al 2008)
By identifying herbs, botanicals and nutritional compounds as well as conventional chemotherapeutic agents that work in synergy with each other our goal is to present to the patient the best options available from both the conventional medical model as well as from and herbal, botanical and nutritional medicine. Please refer to our Comprehensive Cancer Care Consulting Program article to find out more about how we work with cancer.
Understanding Rhabdomyosarcoma
Rhabdomyosarcoma, the most common soft-tissue sarcoma of childhood, results from the neoplastic transformation of cells of the skeletal muscle lineage. Rhabdomyosarcomas harbor a variety of genetic and molecular lesions that frequently include autocrine growth factor circuits along with alterations of oncosuppressor genes. (Nanni et al 2003)
Soft tissue sarcomas are a diverse group of rare tumors that comprise 1% of all cancers. For the majority of histological subtypes adjuvant chemotherapy is not of proven value, although there may be situations where it is advantageous. However, there are other subtypes, such as the Ewing's sarcoma family tumors, for which chemotherapy is an essential part of primary management and has definitely improved survival. Apart from Ewing's sarcoma family tumor and rhabdomyosarcoma, there is increasing specialization of chemotherapy according to histological subtype (Krikelis and Judson 2010)
Soft-tissue sarcomas (STS) include a spectrum of histologically and clinically different tumors. Patients with these tumors are typically relatively young and the course of disease is characterized by early metastasis as well as limited response to chemotherapy. However, a few subtypes, such as small round-cell tumors and rhabdomyosarcoma (other than pleomorphic), are considered chemotherapy sensitive. (Kopp et al 2008)
Rhabdomyosarcoma (RMS) is one of the most common extracranial solid tumours in children. Embryonal and alveolar subtypes of RMS present completely different genetic abnormalities. Embryonal RMS (eRMS) is characterised by loss of heterozygosity on the short arm of chromosome 11 (11p15.5), suggesting inactivation of a tumour-suppressor gene. (Melcon & Codina 2007)
Genomic. Transcription Factor and Growth Factor Tests
1. PDGF
2. C-kit
3. HER II
4. TOPO I & II
5. ERCC-1
6. p27
7. p21
8. p53
9. mdm-2
10. MDR-1
11. PARP-1
12. ras
13. bax
14. Bcl-2
1. IL-8
2. VEGF
3. C-Reactive Protein (CRP)
4. Soluble Transferrin Receptor
5. Vitamin D (OH)
Posible Chemotherapuetic Agents
1. Trabectedin
2. Rapamycin
3. Avastin
4. Low dose Etoposide
Rationale for Genomic, Transcription and Growth Factor Testing
EGFR, PDGFR-alpha, PDGFR-beta, Bcl-2, and Bax are frequently expressed in human rhabdomyosarcoma (RMS) tissue and may represent new therapeutic targets. Absence of PDGFR-alpha and the presence of Bax are associated with a longer median OS in patients with RMS. Targeting these molecules may be a successful therapeutic strategy. (Miyachi et al 2009)
Increased transcriptional levels of PDGF receptors and insulin-like growth factor are associated with decreased survival in rhabdomyosarcomas. Dual blockade of these growth-factor-signaling pathways may be a valuable strategy in preclinical therapeutic studies. (Blandford et al 2006)
PDGF stimulates the proliferation of epithelial cells and other cells. VEGF is related to PDGF and is produced by epithelial cells. (Boik p. 39)
2. C-kit – is a type III receptor tyrosine kinase. It is found that interactions between kit and its ligand, SCF (stem cell factor), are important in the development and maintenance of hematopoietic cells, melanocytes, germ cells, and the interstitial cells. The c-Kit that encodes the receptor for stem cell factor plays a significant role in signal transduction as well as in metastasis. (Bagchi and Preuss 2005) Inhibitors of kit tyrosine activity could have therapeutic efficacy.
The stem cell factor/c-kit pathway may be implicated in the tumorigenesis of spindle cell rhabdomyosarcomaas (RMSs). Therefore, the mutation of c-kit gene must be prospectively examined in larger series of RMSs. (Diniz et al 2006)
HER-2 is expressed in approximately one-half of human rhabdomyosarcomas ; however, its role in the genesis of rhabdomyosarcoma is unknown. Here we show that p53 inactivation coupled with HER-2/neu activation produces rhabdomyosarcomas in mice. (Nanni et al 2003)
4. TOPO I & II - Topoisomerases I & II are enzymes that unwind and wind DNA, in order for DNA to control the synthesis of proteins, and to facilitate DNA replication. The enzyme is necessary due to inherent problems caused by the DNA's double helix.These DNA uncoiling enzymes catalyze the passage of individual DNA strands or double helices through one another. Topo I is most vulnerable to the “S” phase of the cell cycle. In this phase, cells copy their DNA while preparing to divide; therefore, the target enzyme is present more frequently and at a higher concentration.
5. ERCC1 – Excision repair cross-complementing factor 1 - Correlative biomarker studies for near-term trials should include EGFR, ERCC-1, E-cadherin and beta-catenin, epiregulin and amphiregulin, and TP53 mutation. CONCLUSIONS: Revised trial designs in several key areas are needed to advance the study of larynx preservation. With consistent methodologies, clinical trials can more effectively evaluate and quantify the therapeutic benefit of novel treatment options for patients with locally-advanced laryngeal and hypopharyngeal cancer. (Lefebvre and Ang 2009)
6. p27 – is a cyclin-dependent kinase (CDK) inhibitor. Among the beneficial effects the p 27 protein seems to be involved in mediating the growth inhibitory effects of TGF-beta. Loss of p27 activity has been associated with poor prognosis in prostate and colorectal cancer patients. (Boik p. 42-43)
7. p21 - In embryonal rhabdomyosarcomas, p21 may be involved in inhibition of myogenic precursor cell proliferation and differentiation. (Inoue and Wu 2006)
8. p53 – is a transcription factor that controls the expression of genes. It functions to eliminate and inhibit the proliferation of abnormal cells, thereby preventing neoplastic development.
Seventy to eighty percent of rhabdomyosarcoma (RMS) tumors retain wild-type p53. The tumor suppressor p53 plays a central role in inducing cell cycle arrest or apoptosis in response to various stresses. (Miyachi et al 2009)
An increased expression of p53, p21(WAF1), and Bax protein was observed after treatment with MI-63 in rhabdomyosarcoma (RMS) cells with wild-type p53, and apoptosis was confirmed by cleaved PARP and caspase-3 expression. (Canner et al 2009)
The low incidence of rhabdomyosarcoma in individuals carrying p53 alterations indicates that additional genetic lesions are required to cause this tumor. p53 inactivation coupled with HER-2/neu activation produces rhabdomyosarcomas in mice. (Nanni et al 2003)
The highest frequency of p53 mutations reported in human cancers are lung, 56%; colon, 50%; esophagus,45%; ovary, pancreas and skin, 44%; stomach, 41%; head and neck, 37%; bladder, 34%; prostate, 30% and breast, endometrium and mesothelioma, 22%.
NF-κB and p53 pathways together play crucial roles in most human cancers in which hyper-activation of NF-κB and inactivation of p53 is a common occurrence. Inhibition of NF-κB and activation of p53 promotes apoptosis in cancer cells.
9. mdm2 – Further supporting the role of mdm2 as an oncogene, several human tumor types have been shown to have increased levels of mdm2, including soft tissue sarcomas and osteosarcomas as well as breast tumors. (Wikipedia.org)
mdm2 mainly functions to inhibit p 53 activity. In cancer cells, this oncogene is overexpressed, causing chronically low p53 levels and protection from apoptosis. The mdm2 gene suppresses the activity of p53. (Boik 2001 p.18 & 44)
Interruption of the role of p53s as a tumour suppressor by MDM2 may be one of the mechanisms by which cancer cells evade current therapy. (Canner et al 2009)
10. MDR-1- The evaluation of DNA ploidy, p53, MIB1 and MDR-1 expression could be used for subtyping of orbital rhabdomyosarcoma (RMS) into two prognostically different subcategories, respectively RMS responder to the therapy, with favourable clinical outcome, and RMS with a worse prognosis, requiring more aggressive therapeutic protocols. (Staibano et al 2004)
11. PARP-1 - PARP (poly(ADP-ribose)polymerase) – PARP-1 is a nuclear enzyme that is involved in repairing DNA damage (called base excision repair), mediating cell death and necrosis, and regulating immune response. PARP activation occurs when cells are damaged in instances such as during chemotherapy and radiotherapy, and also in non-treatment events such as stroke, head trauma and heart ischemia. The goal of targeting PARP is to prevent tumor cells from repairing DNA themselves and developing drug resistance, which may make them more sensitive to cancer therapies.
Ras induces the expression of the mdm2 gene, whose protein serves to inhibit the activity of the p53 protein. In this way, ras activity reduces the ability of the p53 protein to induce apoptosis in cancer cells. (Boik 2001 p 44)
Oncogenic forms of ras induce several changes in tumor cells, including changes in form and structure and changes in gene expression. The result is excessive DNA synthesis and an increase in chromosomal abnormalities. Ras oncogenes are thought to stimulate the production of enzymes (specifically collagenases) that facilitate tumor invasion. (Boik 2001 p.18)
An increased expression of p53, p21(WAF1), and Bax protein was observed after treatment with MI-63 in Rhabdomyosarcoma (RMS) cells with wild-type p53, and apoptosis was confirmed by cleaved PARP and caspase-3 expression. (Canner et al 2009)
EGFR, PDGFR-alpha, PDGFR-beta, Bcl-2, and Bax are frequently expressed in human rhabdomyosarcoma (RMS) tissue and may represent new therapeutic targets. Absence of PDGFR-alpha and the presence of Bax are associated with a longer median OS in patients with RMS. Targeting these molecules may be a successful therapeutic strategy. (Miyachi et al 2009)
EGFR, PDGFR-alpha, PDGFR-beta, Bcl-2, and Bax are frequently expressed in human rhabdomyosarcoma (RMS) tissue and may represent new therapeutic targets. Absence of PDGFR-alpha and the presence of Bax are associated with a longer median OS in patients with RMS. Targeting these molecules may be a successful therapeutic strategy. (Miyachi et al 2009)
1. IL-8 –. We conclude that IL-8 is a pivotal proangiogenic factor released by human rhabdomyosarcoma (RMS) cells in hypoxic conditions and that the targeting of IL-8 may prove to be a novel and efficient strategy for inhibiting RMS growth. (Wysocozynski et al 2010)
High innate production capacity of proinflammatory cytokines is associated with an increased risk for death from cancer, probably because of increased tumor growth and metastasis. (Trompet et al 2009) IL-8 – is a pro-inflammatory cytokine & an important blood marker to assess angiogenesis/hypoxia. You want it to be low.
2. VEGF – vascular endothelial growth factor – can promote the growth of rhabdomyosarcoma cells through VEGFR1. (Li et al 2006)
Increased levels of VEGF have been associated with many forms of cancers. Because of its role in stimulating angiogenesis near tumors, VEGF is also thought to be an important factor in allowing metastasis to occur. Platelet count and VEGF levels are normally positively correlated. Thus VEGF levels must be corrected when platelets are elevated. Recently, bevacizumab has been approved as an anti-VEGF therapy for metastatic colorectal carcinoma. Laboratory Tests and Diagnostic Procedures, p. 115 ( 2008)
VEGF is one of the most powerful stimulants of tumor angiogenesis. VEGF is a marker of angiogenesis. It is an important blood marker to assess.
VEGF mRNA expression is upregulated by a wide array of oncogenes (including H-ras and K-ras, V-raf, src, PTEN, p53, Wnt, and c-jun, among others) and growth factors (including EGF, TGF-α, TGF-β, insulin-like growth factor-1, and PDGF).
3. CRP – C-reactive Protein is a test that measures the amount of a protein in the blood that signals acute inflammation. The level of CRP rises when there is systemic inflammation. CRP will be up in almost every cancer where the cancer is spreading.
CRP -- is part of the inflammatory process and reflects the level of inflammatory activity. It is associated with inflammation related to insulin resistance. Levels should be below 0.8.
C-Reactive Protein - Because it disappears rapidly when inflammation subsides, its detection signifies the presence of a current inflammatory process. Laboratory Tests and Diagnostic Procedures, p. 393 (2008)
4. Soluble Transferrin Receptor - Our findings show the interest of soluble transferrin receptor and its index in the detection of iron deficiency during anemia of inflammatory states. (Zorgati et al 2009)
Chronic anemia can be due to marrow replacement which may involve tumor cells, fibrous tissue or granulomas. And one of the malignancies that metastasizes to bone marrow resulting in anemia is rhabdomyosarcoma. (From emedicine.medscape.com)
5. Vitamin D is crucial to activating our immune defenses. Without sufficient intake of the vitamin, the killer cells of the immune system -- T cells -- are not be able to react to and fight off serious infections in the body. For T cells to detect and kill foreign pathogens such as clumps of bacteria or viruses, the cells must first be 'triggered' into action and 'transform' from inactive and harmless immune cells into killer cells that are primed to seek out and destroy all traces of a foreign pathogen. It was found that the T cells rely on vitamin D in order to activate and they would remain dormant, 'naïve' to the possibility of threat if vitamin D is lacking in the blood. (von Essen et al 2010)
Moreover, it has been demonstrated that 1,25(OH)(2)D(3) can induce differentiation and inhibit proliferation of normal and malignant cells. Moreover, vitamin D deficiency is associated with an increased risk for nearly all major human diseases such as cancer, autoimmune diseases, cardiovascular, and metabolic diseases. (Verstuyf et al 2010)
Rationale for the Use of Chemotherapeutic Agents
Avastin (Bevacizumab) - Bevacizumab seems to have a good acute safety profile and some antitumor activity in heavily pretreated children and young adults with recurrent solid tumors. (Benesch et al 2008)
Avastin - VEGF can promote the growth of rhabdomyosarcoma cells through VEGFR1, and this effect can be blocked by Avastin. (Li et al 2006)
Rapamycin – Rapamycin induced significant growth inhibition [EFS(T/C) >2] in nine models. Although neither agent given alone caused tumor regressions, in combination, these agents had greater than additive activity against 5 of 13 xenografts and induced complete remissions in one model each of rhabdomyosarcoma and EWS. (Kurmasheva et al 2009)
Rapamycin, a specific inhibitor of mTOR kinase, induces G1 cell cycle arrest and apoptosis in two rhabdomyosarcoma cell lines (Rh1 and Rh30) under conditions of autocrine cell growth. (Hosoi et al 1999) (See section on Understanding mTOR (mammalian target of Rapamycin) for more details on rapamycin and mTOR.)
Etoposide - We prospectively studied the efficacy of high dose therapy (HDT) versus an oral maintenance treatment (OMT) in patients with stage IV soft tissue sarcoma (STS). PATIENTS AND METHODS: Both groups were pretreated with the CEVAIE combination consisting of carboplatin, etoposide, vincristine, actinomycin D, ifosfamide, and epirubicin. HDT consisted of a tandem cycle of thiotepa (600 mg/m(2)) plus cyclophosphamide (4,500 mg/m(2)) and melphalan (120 mg/m(2)) plus etoposide (1,800 mg/m(2)). This treatment was compared with OMT, consisting of four cycles trofosfamide (10 days 2 x 75 mg/m(2)/day) plus etoposide (10 days 2 x 25 mg/m(2)/day), and 4 cycles trofosfamide (10 days 2 x 75 mg/m(2)/day) plus idarubicin (10 days 4 x 5 mg/m(2)). CONCLUSION: Oral maintenance therapy seems to be a promising option for patients with RMS-like stage IV tumors. (Klingebiel et al 2008)
Etoposide - Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children. Current chemotherapy regimes include the topoisomerase II poison etoposide and the transcription inhibitor actinomycin D. CONCLUSIONS: Our findings specifically highlight the topoisomerase poison 9-amino-DACA, its 5-methylsulphone derivative, AS-DACA, and the bis(phenazine-1-carboxamide) transcription inhibitor MLN944/XR5944, currently in phase I trial, as candidates for further research into new agents for the treatment of RMS. (Wolf et al 2009)
Trabectedin - is a tetrahydroisoquinoline molecule that was originally derived from a marine organism. It is indicated in the EU and many other countries for use in patients with advanced soft-tissue sarcoma (STS) who have progressed despite receiving previous treatment with anthracyclines and ifosamide or in those who are unable to receive these agents. Results to date indicate that trabectedin is a valuable addition to the group of second-line antineoplastic agents available for the treatment of advanced, recurrent STS … (Carter & Keam 2010)
Trabectedin has dual effects in liposarcoma: in addition to direct growth inhibition, it affects the tumor microenvironment by reducing the production of key inflammatory mediators. (Germano et al 2010)
Natural Compounds that Target Growth Factors Involved in Cancer
VEGF – vascular endothelial growth factor - is one of the most powerful stimulants of tumor angiogenesis. VEGF mRNA expression is upregulated by a wide array of oncogenes (including H-ras and K-ras, V-raf, src, PTEN, p53, Wnt, and c-jun, among others) and growth factors (including EGF, TGF-α, TGF-β, insulin-like growth factor-1, and PDGF). (Houck et al 1992) (Sima et al 1995) (Shweiki et al 1992) (Toi et al 2001) (Kerbal & Niklinska et al 2001) (Folkman 2002)
VEGF can promote the growth of rhabdomyosarcoma cells through VEGFR1. (Li et al 2006)
Nasal polyp fibroblasts (NPFs) contribute to the pathogenesis of nasal polyps by producing VEGF to promote angiogenesis under hypoxic conditions. Epigallocatechin-3-gallate substantially diminishes HIF-1 alpha and VEGF synthesis in NPFs. (Lin et al 2008)
Natural Compounds that Inhibit VEGF:
Artemisia annua (Chen et al 2004)
Camellia sinensis (green tea extract) (Lin et al 2008)
Curcuma longa (turmeric) (Schaaf et al 2010)
Ginkgo biloba (Zhang et al 2002)
Magnolia seed cones – 90% honokiol (Wen et al 2009) (Ahn et al 2006)
Ocimum spp. (Basil) Ursolic acid (Yan et al 2010)
Polygonum cuspidatum (Japanese knotweed) (Wang et al 2004) (Cao et al 2005) – 20% resveratrol
Pterostilbene – (Pan et al 2009)
Scutellaria baicalensis (Yoon et al 2009)
Silybum marianum (milk thistle) (Gallo et al 2003)
Taxus breviflora (Pacific yew) – taxol and other related taxins
Viscum album (Mistletoe) (Park et al 2001)
Vitus vinifera (grape seed extract) (Bai et al 2003)
p53 – the Guardian of the Genome.
The highest frequency of p53 mutations reported in human cancers are lung, 56%; colon, 50%; esophagus,45%; ovary, pancreas and skin, 44%; stomach, 41%; head and neck, 37%; bladder, 34%; prostate, 30% and breast, endometrium and mesothelioma, 22%.
p53 restoration therapy is a potential therapeutic strategy for refractory rhabdomyosarcoma with wild-type p53. (Miyachi et al 2009)
Natural Compounds that Normalize p53
6-Gingerol (Park et al 2006)
Coriolus versicolor (Ho et al 2005)
Curcumin (Boik 2001 p.55)
EGCG (Lee et al 2010)
Tocotrienols (Agarwal et al 2004) and Vit E succinate
OPCs
Oridonin (Rabdosia) (Cui et al 2007)
Paw paw seed
Quercetin (Beniston et al 2001)(Shi et al 2003)
Resveratrol (She et al 2001)(Pearce et al 2007)(Tang et al 2006)
Rhemannia glutinosa (Chao et al 2006)
Withanone (Ashwagandha extract) (Widodo et al 2008)
Note: Diets rich in refined sugars and starches, as well as rich in red meat promote p53 mutation. (Slattery et al 2002)
Bcl-2 – is an NF-κB regulated gene that functions by blocking the apoptosis pathway, thus immortalizing cancer cells. It has been suggested that Bcl-2 over expression results in the up regulation of VEGF expression with increased neoangiogenesis in human cancer xenografts.
Natural Compounds that Reduce Bcl-2
Andrographolide (Zhou et al 2006) – Andrographis
Baicalin and baicalein (Chen et al 2000) - Scutellaria baicalensis
Beta-lapachone – Lapacho (Woo et al 2005)
Carnosol – Rosemary (Dorrie et al 2001)
Casticin – Yarrow (Achillea millefolium) (Khadidja et al 2006)
Chelidonium alkaloids – Chelidonine (Habermehl et al 2006)
Curcumin (Bharti et al 2003) (Schaaf et al 2010)
DIM – (Hong et al 2002)
Echinocystic acid (EA) (Tong et al 2004) – Panax ginseng
EGCG (Lee et al 2010) and Theophylline - Green tea (Leone et al 2003)(Byrd et al 2000)
EPA – from fish oil (Chiu and Wan 1999)
Gingerol – Ginger (Wang et al 2003)
Hibiscus protocatechuic acid (PCA) (Tseng et al 2000)
Mistletoe – (Choi et al 2004)
OPCs – grape seed extract – (Bagchi et al 2000)
Parthenolide – feverfew (Zhang et al 2004)
Resveratrol – (Aziz et al 2006)
NF-kappaB - is an important transcription factor that is up-regulated in practically all cancers. It up regulates inflammation, angiogenesis, metastasis and tumor promotion. It is hyperactive in many cancers raising the resistance of cancer cells to chemotherapy drugs and chemo radiation.
Natural Compounds that Inhibit or Down-Regulate NF-kappaB:
Chinese skullcap (Piao et al 2008) (Peng et al 2008)
Curcumin – (Rafiee et al 2010) (Kamat et al 2009)
DIM (Banerjee et al 2009)(Rahman et al 2007) and I-3-C (Rahman et al 2006) (Takada et al 2005)
Green Tea Polyphenols EGCG (Lee et al 2009) (SBabu et al 2009) (Siddiqui et al 2008)
Magnolol (Ahn et al 2006)
N-acetyl cysteine (NAC) (Sailai et al 2010)
Panax ginseng (Jou et al 2009) and panax notoginseng (Son et al 2009)
Parthenolide (Nehra et al 2010) (Riggins et al 2005)
Pterostilbene (Pan et al 2009) and quercetin (Granado-Serrano et al 2010)
Reishi powdered extract ((Jiang et al 2008) (Ho et al 2007)
Resveratrol (Singh et al 2010) (Park et al 2009) (Benitez et al 2009)
Silymarin (Momeny et al 2010)
Sulforaphane (PEITC) (Xu et al 2005) (Brunelli et al 2010)
Ursolic acid (Manu & Kuttan 2008) – in Holy Basil
COX-2 is up-regulated in practically all cancers. It is induced by phorbol esters, cytokines and growth factors, including TGF-beta-1 and bFGF. COX-2 is a potent inducer of angiogenesis by inducing angiogenic factors. Most common cancers with altered (amplified) COX-2 expression include: prostate, colon, breast, cervical brain, gastric, pancreatic, lung, head and neck, kidney and bladder.
Natural Compounds that Inhibit or Down-Regulate COX-2:
EPA and DHA in n-3 fatty acids from fish oils (Lee et al 2009) (Lim et al 2009)
Baicalein, from Chinese skullcap (Chiu et al 2010)
Curcumin (Lin et al 2010) (Moon et al 2010) (Leite et al 2009)
Resveratrol (Kang et al 2009)
Panax notoginseng (Son et al 2009)
Parthenolide (Weng et al 2009)
Pterostilbene (Pan et al 2008)
Quercetin (Lee et al 2010) (Turner et al 2009) (Warren et al 2009)
Salicin (Corydalis 30%) Willow bark extract (WBE)
WBE inhibits the cell growth and promote apoptosis in human colon and lung cancer both through COX-selectivity and nonCOX-2. Other synergistic compound WBE include other salicyl alcohol derivates, flavonoids, proanthocyanidins.
Several foods and common culinary herbs contain particular compounds that reduce inflammation, in part, by acting as COX-2 inhibitors. These include:
Grapes (leaf and skin)
Curcumin longo (turmeric)
Ocimun sanctum (basil)
Fish
Flax
Oregano
Rosmarinus (rosemary)
Licorice root (glycyrrhizin glabra)
PDGF - platelet-derived growth factor – Increased transcriptional levels of PDGF receptors and insulin-like growth factor are associated with decreased survival in rhabdomyosarcomas. (Blandford et al 2006)
Expression of PDGF, and activation of its receptor (PDGFR), a tyrosine kinase, is associated with the growth of metastasis in several forms of cancer including kidney, breast, prostate, sarcomas, cervical and in head and neck cancer. Inhibition of PDGF has shown to suppress metastatic cancer.
Natural Compounds that Suppress PDGF
Curcumin causes an interruption of the PDGF and EGF signaling pathways by stimulating gene expression of PPARγ.
Curcumin, also decreases the proportion of S phase cells after PDGF stimulation
Baicalein, in Chinese skullcap, one of the most powerful anti-cancer agents induces apoptosis of cancer cells. One known mechanism is by down-regulating 12-LOX – reducing PDGF.
EGCG, from GTE, inhibits PDGF-induced VEGF expression via blocking PDGF receptor and Erk-1/2.
Understanding Transcription Factors
Transcription factors are proteins that bind to specific DNA sequences and thereby control the transfer (or transcription) of genetic information from DNA to mRNA. They are essential for the regulation of gene expression. There are approximately 2600 proteins in the human genome that contain DNA-binding domains and most of these are presumed to function as transcription factors. They use a variety of mechanisms for the regulation of gene expression such as: 1) Histone acetyltransferease (HAT) activity – this acetylates histone proteins, which weakens the association of DNA histones, which make the DNA more accessible to transcription and thereby upregulate transcription and 2) Histone deacetylase (HDAC) activity. This deacetylates histone proteins, which strengthens the association of DNA histones, which make the DNA less accessible to transcription and thereby down regulate transcription.
Not only do transcription factors act downstream of signaling cascades related to biological stimuli but they can also be downstream of signaling cascades involved in environmental stimuli. Examples include heat shock factor (HSF), which up regulates genes necessary for survival at higher temperatures and hypoxia inducible factor (HIF), which upregulates genes necessary for cell survival in low-oxygen environments. Examples of transcription factors include: p53, HSF1, and NF-kappaB. (Answers.com Genetics Encylopedia)
Diagram Showing Various Transcription and Signaling Pathways in the Cell such as: mdm2, p53, ras, mTOR, NF-kappaB, etc. (Click on Page if Blank)
The location and role of mTOR in the cell.
o mTOR is a key intracellular point of convergence for a number of cellular signaling pathways. mTOR performs its regulatory function in response to activating or inhibitory signals transmitted through these pathways, which are located upstream from mTOR in the cell. These diverse signaling pathways are activated by a variety of growth factors (including vascular endothelial growth factors (VEGFs), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin-like growth factor 1 (IGF-1)), hormones (estrogen, progesterone), and the presence or absence of nutrients (glucose, amino acids) or oxygen.4,5
Abstracts Cited in this Article
Expert Rev Anticancer Ther. 2010 Feb;10(2):249-60.
University Hospital of Ioannina, Stavros Niarchos Avenue, 45500, Ioannina, Greece. krikelis@yahoo.com
Soft tissue sarcomas are a diverse group of rare tumors that comprise 1% of all cancers. Few randomized trials of chemotherapy have been performed but there is a clear role for agents such as doxorubicin and ifosfamide in the palliation of advanced disease. There is uncertainty as to whether sequential single-agent treatment is equivalent to combination chemotherapy. For the majority of histological subtypes adjuvant chemotherapy is not of proven value, although there may be situations where it is advantageous. However, there are other subtypes, such as the Ewing's sarcoma family tumors, for which chemotherapy is an essential part of primary management and has definitely improved survival. Apart from Ewing's sarcoma family tumor and rhabdomyosarcoma, there is increasing specialization of chemotherapy according to histological subtype, such as the use of taxanes for angiosarcoma, gemcitabine and docetaxel for leiomyosarcoma, and trabectedin for leiomyosarcoma and liposarcoma, especially the myxoid/round cell variant. Nevertheless, there are serious limitations to existing treatment and novel therapies need to be developed.
Am J Clin Dermatol. 2008;9(4):207-17.
Kopp HG, Patel S, Brücher B, Hartmann JT.
Medical Center II, Department of Oncology, Hematology, Rheumatology, Immunology, and Pulmonology, South West German Comprehensive Cancer Center, Eberhard-Karls-University Tuebingen, Tuebingen, Germany.
Soft-tissue sarcomas (STS) include a spectrum of histologically and clinically different tumors. Patients with these tumors are typically relatively young and the course of disease is characterized by early metastasis as well as limited response to chemotherapy. However, a few subtypes, such as small round-cell tumors and rhabdomyosarcoma (other than pleomorphic), are considered chemotherapy sensitive. In addition, reflecting successful translational research of recent years, gastrointestinal stromal tumor and dermatofibrosarcoma protuberans have become model diseases for targeted oncologic therapy. We summarize current treatment options for metastatic STS, including established first-line chemotherapy approaches, mainly with anthracyclines and/or ifosfamide and second-line treatment choices beyond anthracyclines. Until only a few years ago, treatment choices for metastatic STS were easy to review because of the very limited number of active compounds available. However, with the advent of novel therapeutic strategies such as the anti-angiogenic approach and a multitude of novel compounds available both outside and within clinical studies, it has potentially become more difficult to keep track of currently available treatment options for STS and their clinical safety and efficacy. In this practice-oriented article, we therefore review treatment goals in advanced STS and provide an overview of compounds with proven activity in this setting. Anthracyclines with or without ifosfamide are still considered standard of care for most STS subtypes, especially for high-grade tumors. There is no evidence-based recommendation regarding use of second-line treatment options. However, a number of established compounds, including dacarbazine/temozolomide, gemcitabine, taxanes, trofosfamide, DNA topoisomerase I inhibitors, DNA minor groove binders, and bendamustine have shown activity. Recently, trabectedin, a DNA minor groove binder initially isolated from a sea sponge, has proven effective and received European approval for use in treatment-refractory STS. In addition, novel compounds such as bevacizumab, multi-tyrosine kinase inhibitors, mammalian target of rapamycin inhibitors, imatinib, and the thrombospondin agonist ABT 510 represent attractive partners for the above-mentioned cytostatic agents, or may even be effective single agents in the clinically advanced setting. Novel combinations are being evaluated in clinical studies. In order to be successful, it may be necessary to combine not only different compounds but also different targets beyond the proliferation machinery of sarcoma cells such as tumor angiogenesis, the tumor stromal compartment, or tumor cell oncogene products.
Ann Oncol. 2008 Apr;19(4):807-13. Epub 2007 Dec 3.
Benesch M, Windelberg M, Sauseng W, Witt V, Fleischhack G, Lackner H, Gadner H, Bode U, Urban C.
Division of Pediatric Hematology and Oncology, Department of Pediatrics and Adolescent Medicine, Medical University of Graz, Austria. martin.benesch@klinikum-graz.at
BACKGROUND: The aim of this study was to evaluate feasibility and toxicity of bevacizumab (Avastin), a monoclonal antibody directed against the vascular endothelial growth factor in children and young adults. PATIENTS AND METHODS: Fifteen patients (male: n = 8; female: n = 7; median age, 14.6 years) received bevacizumab for recurrent or progressive solid tumors (carcinoma: n = 3; neuroblastoma: n = 2; astrocytoma grade III: n = 2; rhabdomyosarcoma: n = 2; nephroblastoma: n = 2; benign vascular tumors: n = 2; synovial sarcoma: n = 1; and malignant hemangiopericytoma: n = 1) on a compassionate basis. Bevacizumab was administered at 5-10 mg/kg body weight intravenously every 2-3 weeks. Most patients received chemotherapy in addition to bevacizumab. Duration of bevacizumab therapy ranged from 1.5 to 23 months. RESULTS: Bevacizumab-related side-effects were mild and included hypertonia (n = 2), proteinuria/hematuria (n = 2), epistaxis (n = 2), local erythema (n = 1), and defective wound healing and ascites (n = 1). Radiographic objective responses (partial responses) were observed in two patients with astrocytoma grade III and in one patient each with neuroblastoma and pleomorphic rhabdomyosarcoma, respectively. CONCLUSIONS: Bevacizumab seems to have a good acute safety profile and some antitumor activity in heavily pretreated children and young adults with recurrent solid tumors. Prospective clinical trials are urgently needed to further evaluate the safety and efficacy of bevacizumab in pediatric patients.
Cancer Res. 2009 Oct 1;69(19):7662-71. Epub 2009 Sep 29.
Kurmasheva RT, Dudkin L, Billups C, Debelenko LV, Morton CL, Houghton PJ.
Departments of Molecular Pharmacology, Biostatistics, and Pathology, St. Jude Children's Research Hospital, Memphis, TN38105, USA.
Signaling through the type 1 insulin-like growth factor receptor (IGF-1R) occurs in many human cancers, including childhood sarcomas. As a consequence, targeting the IGF-1R has become a focus for cancer drug development. We examined the antitumor activity of CP-751,871, a human antibody that blocks IGF-1R ligand binding, alone and in combination with rapamycin against sarcoma cell lines in vitro and xenograft models in vivo. In Ewing sarcoma (EWS) cell lines, CP751,871 inhibited growth poorly (<50%), but prevented rapamycin-induced hyperphosphorylation of AKT(Ser473) and induced greater than additive apoptosis. Rapamycin treatment also increased secretion of IGF-1 resulting in phosphorylation of IGF-1R (Tyr1131) that was blocked by CP751,871. In vivo CP-751,871, rapamycin, or the combination were evaluated against EWS, osteosarcoma, and rhabdomyosarcoma xenografts. CP751871 induced significant growth inhibition [EFS(T/C) >2] in four models. Rapamycin induced significant growth inhibition [EFS(T/C) >2] in nine models. Although neither agent given alone caused tumor regressions, in combination, these agents had greater than additive activity against 5 of 13 xenografts and induced complete remissions in one model each of rhabdomyosarcoma and EWS, and in three of four osteosarcoma models. CP751,871 caused complete IGF-1R down-regulation, suppression of AKT phosphorylation, and dramatically suppressed tumor-derived vascular endothelial growth factor (VEGF) in some sarcoma xenografts. Rapamycin treatment did not markedly suppress VEGF in tumors and synergized only in tumor lines where VEGF was dramatically inhibited by CP751,871. These data suggest a model in which blockade of IGF-1R suppresses tumor-derived VEGF to a level where rapamycin can effectively suppress the response in vascular endothelial cells.
Pediatr Blood Cancer. 2008 Apr;50(4):739-45.
Klingebiel T, Boos J, Beske F, Hallmen E, Int-Veen C, Dantonello T, Treuner J, Gadner H, Marky I, Kazanowska B, Koscielniak E.
Children's Hospital of Frankfurt University Hospital, Frankfurt, Germany. thomas.klingebiel@kgu.de
PURPOSE: We prospectively studied the efficacy of high dose therapy (HDT) versus an oral maintenance treatment (OMT) in patients with stage IV soft tissue sarcoma (STS). PATIENTS AND METHODS: Both groups were pretreated with the CEVAIE combination consisting of carboplatin, etoposide, vincristine, actinomycin D, ifosfamide, and epirubicin. HDT consisted of a tandem cycle of thiotepa (600 mg/m(2)) plus cyclophosphamide (4,500 mg/m(2)) and melphalan (120 mg/m(2)) plus etoposide (1,800 mg/m(2)). This treatment was compared with OMT, consisting of four cycles trofosfamide (10 days 2 x 75 mg/m(2)/day) plus etoposide (10 days 2 x 25 mg/m(2)/day), and 4 cycles trofosfamide (10 days 2 x 75 mg/m(2)/day) plus idarubicin (10 days 4 x 5 mg/m(2)). Eligibility criteria were: diagnosis confirmed by reference pathology, primary stage IV, below 22 years of age, and having completed the study therapy. RESULTS: From 96 patients 45 were treated with HDT and 51 with OMT. The main risk parameters were equally distributed in both arms. After a median follow-up of 57.4 months, 11/45 (24.4%) patients in the HDT-arm and 26/51 (57.8%) patients in OMT-arm were alive. Kaplan-Meier analysis demonstrated an overall survival for the whole group of 0.27 (OMT group: 0.52, HDT group 0.27, log rank P = 0.03). The proportional hazard analysis for patients with rhabdomyosarcoma (RMS) or "RMS-like" tumors (77.1% of all patients) demonstrated an independent benefit of OMT on outcome. CONCLUSION: Oral maintenance therapy seems to be a promising option for patients with RMS-like stage IV tumors.
Cancer Chemother Pharmacol. 2009 Nov;64(6):1059-69. Epub 2009 Mar 10.
Wolf SJ, Wakelin LP, He Z, Stewart BW, Catchpoole DR.
The Tumour Bank, Oncology Research Unit, The Children's Hospital at Westmead, Locked Bag 4001, Westmead, NSW 2145, Australia.
PURPOSE: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children. Current chemotherapy regimes include the topoisomerase II poison etoposide and the transcription inhibitor actinomycin D. Poor clinical response necessitate identification of new agents to improve patient outcomes. METHODS: We assessed the in vitro cytotoxicity (MTT assay) of DNA intercalating agents in five established human RMS cell lines. These include novel classes of transcription inhibitors and topoisomerase poisons, previously shown to have potential as anti-cancer agents. RESULTS: Amongst the former agents, bisintercalating bis(9-aminoacridine-4-carboxamides) linked through the 9-position, and bis(phenazine-1-carboxamides) linked via their side chains, are compared with established transcription inhibitors. Amongst the latter, monofunctional acridine-4-carboxamides related to N-[2-(dimethylamino)ethyl]acridine-4-carboxamide, DACA, are compared with established topoisomerase poisons. CONCLUSIONS: Our findings specifically highlight the topoisomerase poison 9-amino-DACA, its 5-methylsulphone derivative, AS-DACA, and the bis(phenazine-1-carboxamide) transcription inhibitor MLN944/XR5944, currently in phase I trial, as candidates for further research into new agents for the treatment of RMS.
Ai Zheng. 2006 Jun;25(6):718-22.
[Article in Chinese]
Li ZG, Shi YX, Peng RJ, Zhu MQ, Wan YY.
School of Basic Medicine, Sun Yat-sen University, Guangzhou, Guangdong, 510080, P. R. China.
BACKGROUND & OBJECTIVE: Vascular endothelial growth factor (VEGF) signal pathway is a hot spot in recent tumor research, but its role in rhabdomyosarcoma (RMS) has rarely been reported, and its mechanism is unclear. This study was to investigate the expression of VEGF and its receptors (VEGFR) in RMS cell line RH4 to reveal the mechanism of VEGF signal pathway, and explore the therapeutic value of Avastin in RMS in vitro. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of VEGF, VEGFR1, VEGFR2, and VEGFR3 mRNA in RH4 cells. ELISA was used to detect the concentration of VEGF in supernatant. Western blot was used to detect the expression of VEGFR1 protein. The effects of VEGF and Avastin on RH4 cell growth were evaluated by direct cell counting. RESULTS: RH4 cells expressed VEGF at a high level and secreted VEGF to culture media. RH4 cells only expressed VEGFR1, did not express VEGFR2 and VEGFR3. VEGF promoted the growth of RH4 cells in a dose-dependent manner within a concentration range of 0-100 ng/ml, this effect could be blocked by 10-40 microg/ml Avastin. CONCLUSION: VEGF can promote the growth of RMS cell through VEGFR1, and this effect can be blocked by Avastin.
Drugs. 2010 Feb 12;70(3):355-76. doi: 10.2165/11202860-000000000-00000.
Adis, a Wolters Kluwer Business, Auckland, New Zealand. demail@adis.co.nz
Trabectedin (Yondelis) is a tetrahydroisoquinoline molecule that was originally derived from a marine organism. It is indicated in the EU and many other countries for use in patients with advanced soft-tissue sarcoma (STS) who have progressed despite receiving previous treatment with anthracyclines and ifosfamide or in those who are unable to receive these agents. It is also approved in the EU in combination with pegylated liposomal doxorubicin for the treatment of platinum-sensitive, recurrent ovarian cancer. In addition, trabectedin holds orphan drug status for the treatment of advanced, recurrent STS in the US, Switzerland and Korea, and for the treatment of advanced, recurrent ovarian cancer in the US and Switzerland. Clinical trials showed that intravenous trabectedin was effective in chemotherapy-experienced patients with advanced, recurrent liposarcoma or leiomyosarcoma, and results from a retrospective analysis suggest that the drug may be particularly effective in patients with advanced myxoid liposarcoma. In addition, coadministration of trabectedin with pegylated liposomal doxorubicin was associated with a significantly longer progression-free survival (6 weeks) than pegylated liposomal doxorubicin monotherapy in patients with recurrent ovarian cancer after failure of first-line, platinum-based chemotherapy. The tolerability profile of trabectedin was manageable in clinical trials, and the tolerability profile of concomitant trabectedin and pegylated liposomal doxorubicin was generally consistent with that of each agent alone. Results to date indicate that trabectedin is a valuable addition to the group of second-line antineoplastic agents available for the treatment of advanced, recurrent STS, and that it is a beneficial treatment for recurrent ovarian cancer after failure of first-line, platinum-based chemotherapy when administered in conjunction with pegylated liposomal doxorubicin.
Cancer Res. 2010 Mar 15;70(6):2235-44. Epub 2010 Mar 9.
Germano G, Frapolli R, Simone M, Tavecchio M, Erba E, Pesce S, Pasqualini F, Grosso F, Sanfilippo R, Casali PG, Gronchi A, Virdis E, Tarantino E, Pilotti S, Greco A, Nebuloni M, Galmarini CM, Tercero JC, Mantovani A, D'Incalci M, Allavena P.
Department of Immunology and Inflammation, IRCCS Istituto Clinico Humanitas, Rozzano, Italy.
Inflammatory mediators present in the tumor milieu may promote cancer progression and are considered promising targets of novel biological therapies. We previously reported that the marine antitumor agent trabectedin, approved in Europe in 2007 for soft tissue sarcomas and in 2009 for ovarian cancer, was able to downmodulate the production of selected cytokines/chemokines in immune cells. Patients with myxoid liposarcoma (MLS), a subtype characterized by the expression of the oncogenic transcript FUS-CHOP, are highly responsive to trabectedin. The drug had marked antiproliferative effects on MLS cell lines at low nanomolar concentrations. We tested the hypothesis that trabectedin could also affect the inflammatory mediators produced by cancer cells. Here, we show that MLS express several cytokines, chemokines, and growth factors (CCL2, CCL3, CCL5, CXCL8, CXCL12, MIF, VEGF, SPARC) and the inflammatory and matrix-binder protein pentraxin 3 (PTX3), which build up a prominent inflammatory environment. In vitro treatment with noncytotoxic concentrations of trabectedin selectively inhibited the production of CCL2, CXCL8, IL-6, VEGF, and PTX3 by MLS primary tumor cultures and/or cell lines. A xenograft mouse model of human MLS showed marked reduction of CCL2, CXCL8, CD68+ infiltrating macrophages, CD31+ tumor vessels, and partial decrease of PTX3 after trabectedin treatment. Similar findings were observed in a patient tumor sample excised after several cycles of therapy, indicating that the results observed in vitro might have in vivo relevance. In conclusion, trabectedin has dual effects in liposarcoma: in addition to direct growth inhibition, it affects the tumor microenvironment by reducing the production of key inflammatory mediators.
Pediatr Blood Cancer. 2006 Mar;46(3):329-38.
Blandford MC, Barr FG, Lynch JC, Randall RL, Qualman SJ, Keller C.
Department of Pediatrics, Division of Pediatric Hematology-Oncology, University of Utah, Salt Lake City, USA.
BACKGROUND: Unresectable or metastatic disease represents the greatest obstacle to cure for children with rhabdomyosarcoma. In this study we sought to identify gene expression signatures of advanced stage and progressive disease. PROCEDURE: Using oligonucleotide gene expression analysis for a focused set of 60 genes, we analyzed the myogenic expression profiles of 89 rhabdomyosarcomas from the Intergroup Rhabdomyosarcoma Study-IV. RESULTS: While the expression profile of rhabdomyosarcomas closely paralleled gene expression profiles of normal embryonic myogenic progenitors, growth factors were most closely associated with disease progression. Specifically, we identified platelet-derived growth factor (PDGF) receptors and insulin-like growth factor as strongly correlated with decreased failure-free survival. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) of an independent data set suggested that autocrine growth signaling, if present, is not regulated in a simple manner at the transcriptional level. CONCLUSIONS: Increased transcriptional levels of PDGF receptors and insulin-like growth factor are associated with decreased survival in rhabdomyosarcomas. Dual blockade of these growth-factor-signaling pathways may be a valuable strategy in preclinical therapeutic studies.
Pathol Res Pract. 2006;202(9):671-7. Epub 2006 Jul 24.
Diniz G, Aktas S, Ortac R, Tunakan M, Unlu I, Vergin C.
Pathology and Pediatric Oncology/Hematology Department, Izmir Dr.Behcet Uz Children's Hospital, Turkey. agdiniz@kablonet.com.tr
The use of a relatively nontoxic tyrosine kinase receptor inhibitor, imatinib mesylate (IM) (STI-571), has increasingly become a valuable therapeutic alternative in some KIT (CD117)-overexpressing neoplasms potentially because of the presence of KIT-activating mutations. As the treatment eligibility for this drug hinges on CD117 expression, KIT immunostaining has recently been widely examined in various different tumors. We examined CD117 expression in pediatric embryonal rhabdomyosarcomas (RMSs) to identify its eventual prognostic impact and to evaluate its effect on tumorigenesis. This study included two spindle cell (leiomyomatous) variants, two botryoid variants, and 21 conventional embryonal RMSs. Sections from paraffin-embedded tumor samples were immunostained by a standard SABC technique using c-kit polyclonal antibody with antigen retrieval. In all the series, the percentage of CD117 positivity was 12%. Staining was strong in two of two spindle cell variants, in zero of two botryoid variants, and in one of 21 conventional embryonal RMSs. In Spearman's correlation analysis, there was statistical relationship between the presence of CD117 expression and the histological subtype of RMS. Kaplan-Meier analysis revealed no prognostic significance of CD117 expression for survival. The present study demonstrated a very limited expression of CD117 in pediatric embryonal RMS other than in the spindle cell variant. This finding suggested that the stem cell factor/c-kit pathway may be implicated in the tumorigenesis of spindle cell RMSs. Therefore, the mutation of c-kit gene must be prospectively examined in larger series of RMSs. If it can be verified that tissue expression of CD117 reflects the mutation of c-kit gene, IM can be considered a targeted therapy for CD117-expressing RMSs, particularly the spindle cell variant.
Tunis Med. 2009 Dec;87(12):818-23.
[Article in French]
Zorgati MM, Hafsia R, Bahlous A, Bahri S, Ben Ammar S.
Laboratoire Central de Biologie Médicale, Institut Pasteur de Tunis.
BACKGROUND: In inflammatory diseases, classical parameters of iron status (serum iron, serum ferritin, total iron-binding capacity of transferrin and transferrin saturation) are not very reliable. AIM: The purpose of this study is to investigate soluble transferrin receptor, its index and classical parameters of iron status [serum iron, serum ferritin, total iron-binding capacity of transferrin and transferrin saturation] during iron-deficiency anemia and combined iron deficiency and inflammatory anemia. METHODS: Our study concerned 24 patients: 18 patients with iron-deficiency anemia and 6 patients with combined iron-deficiency and inflammatory anemia. 55 healthy subjects were included as controls. Both groups underwent classical parameters of iron status [serum iron, serum ferritin, total iron-binding capacity of transferrin and transferrin saturation] and measurement of soluble transferrin receptor with its index. RESULTS: In iron-deficiency anemia, total iron-binding capacity of transferrin, soluble transferrin receptor and its index were enhanced, whereas serum iron, ferritinemia and transferrin saturation were low compared to controls. Compared to patients with iron-deficiency anemia, those with combined iron-deficiency and inflammatory anemia showed higher levels of serum iron and ferritinemia. In contrast, soluble transferrin receptor and its index did not vary significantly between both groups. CONCLUSION: Our findings show the interest of soluble transferrin receptor and its index in the detection of iron deficiency during anemia of inflammatory states.
Am J Pathol. 2009 Feb;174(2):550-64. Epub 2009 Jan 15.
Davicioni E, Anderson MJ, Finckenstein FG, Lynch JC, Qualman SJ, Shimada H, Schofield DE, Buckley JD, Meyer WH, Sorensen PH, Triche TJ.
Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA.
Rhabdomyosarcoma (RMS) in children occurs as two major histological subtypes, embryonal (ERMS) and alveolar (ARMS). ERMS is associated with an 11p15.5 loss of heterozygosity (LOH) and may be confused with nonmyogenic, non-RMS soft tissue sarcomas. ARMS expresses the product of a genomic translocation that fuses FOXO1 (FKHR) with either PAX3 or PAX7 (P-F); however, at least 25% of cases lack these translocations. Here, we describe a genomic-based classification scheme that is derived from the combined gene expression profiling and LOH analysis of 160 cases of RMS and non-RMS soft tissue sarcomas that is at variance with conventional histopathological schemes. We found that gene expression profiles and patterns of LOH of ARMS cases lacking P-F translocations are indistinguishable from conventional ERMS cases. A subset of tumors that has been histologically classified as RMS lack myogenic gene expression. However, classification based on gene expression is possible using as few as five genes with an estimated error rate of less than 5%. Using immunohistochemistry, we characterized two markers, HMGA2 and TFAP2ss, which facilitate the differential diagnoses of ERMS and P-F RMS, respectively, using clinical material. These objectively derived molecular classes are based solely on genomic analysis at the time of diagnosis and are highly reproducible. Adoption of these molecular criteria may offer a more clinically relevant diagnostic scheme, thus potentially improving patient management and therapeutic RMS outcomes.
Genes Chromosomes Cancer. 2009 Nov;48(11):975-82.
Martinelli S, McDowell HP, Vigne SD, Kokai G, Uccini S, Tartaglia M, Dominici C.
Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy. simone.martinelli@iss.it
Rhabdomyosarcoma (RMS) is a common childhood solid tumor, resulting from dysregulation of the skeletal myogenesis program. Two major histological subtypes occur in childhood RMS, embryonal and alveolar. While chromosomal rearrangements account for the majority of alveolar tumors, the genetic defects underlying the pathogenesis of embryonal RMS remain largely undetermined. A few studies performed on small series of embryonal tumors suggest that dysregulation of RAS function may be relevant to disease pathogenesis. To explore further the biological and clinical relevance of mutations with perturbing consequences on RAS signaling in embryonal RMS, we investigated the prevalence of PTPN11, HRAS, KRAS, NRAS, BRAF, MEK1, and MEK2 mutations in a relatively large cohort of primary tumors. While HRAS and KRAS were found to be rarely mutated, we identified somatic NRAS lesions in 20% of cases. All mutations were missense and affected codon 61, with the introduction of a positive charged amino acid residue representing the most common event. PTPN11 was found mutated in one tumor specimen, confirming that somatic defects in this gene are relatively uncommon in RMS, while no mutation was observed in BRAF and MEK genes. Although no clear association of mutations with any clinical variable was observed, comparison of the outcome between mutation-positive and mutation-negative cases indicated a trend for a higher percentage of patients exhibiting a better outcome in the former. Our findings provide evidence that dysregulation of RAS signaling is a major event contributing to embryonal RMS pathogenesis.
Int J Cancer. 2010 Jan 15;126(2):371-81.
Wysoczynski M, Shin DM, Kucia M, Ratajczak MZ.
James Graham Brown Cancer Center, University of Louisville, KY 40202, USA.
Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma of adolescence and childhood. Because RMS tumors are highly vascularized, we sought to determine which factors secreted by RMS cells are crucial in stimulating angiogenesis in response to hypoxia. To address this issue, we evaluated expression of several proangiogenic factors [interleukin (IL)-8, vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF)-2, stromal-derived factor (SDF)-1, hepatocyte growth factor (HGF) and leukemia inhibitory factor (LIF)] in 8 human RMS cell lines in both normal steady-state and hypoxic conditions. We found by real-time quantitative polymerase chain reaction (RQ-PCR) and confirmed by enzyme-linked immunosorbent assay (ELISA) that from all the factors evaluated, IL-8, whose expression is very low in normoxia, had been very highly expressed and secreted by RMS cells lines during hypoxic conditions ( approximately 40-170 times). Interestingly, this upregulation was not affected by knocking down hypoxia-inducible factor (HIF)-1alpha, but was inhibited by mitogen-activated protein kinase (MAPK)p42/44 and phosphatidylinositaol 3-kinase (PI3K)/AKT pathway inhibitors. This suggests that IL-8 expression is regulated in an activating protein (AP)-1- and nuclear factor (NF)-kappaB-dependent manner. Furthermore, we found that conditioned media (CM) harvested from RMS cells exposed to hypoxia activated and stimulated chemotactic responses in human umbilical vein endothelial cells (HUVECs) and that IL-8 was responsible for hypoxia-related effects. Finally, by employing shRNA, the expression of IL-8 in human RH-30 cells was downregulated. We noticed that such RMS cells, if injected into skeletal muscles of immunodeficient mice, have a reduced ability for tumor formation. We conclude that IL-8 is a pivotal proangiogenic factor released by human RMS cells in hypoxic conditions and that the targeting of IL-8 may prove to be a novel and efficient strategy for inhibiting RMS growth.
Clin Cancer Res. 2009 Dec 15;15(24):7744-7748.
Trompet S, de Craen AJ, Mooijaart S, Stott DJ, Ford I, Sattar N, Jukema W, Westendorp RG.
Authors' Affiliations: Departments of Gerontology and Geriatrics, and Cardiology, Leiden University Medical Center, Leiden, the Netherlands; Durrer Center for Cardiogenetic Research, Amsterdam, the Netherlands; Department of Geriatric Medicine, Robertson Centre for Biostatistics, and British Heart Foundation Glasgow Cardiovascular Research Centre, Faculty of Medicine, University of Glasgow, Glasgow, Scotland; and Netherlands Consortium of Healthy Ageing, Leiden, the Netherlands.
PURPOSE: Various lines of evidence suggest that proinflammatory factors may play a role in tumor growth and metastasis, the leading cause of cancer-related mortality. However, most evidence originates from animal models, only few human studies reported an association between proinflammatory cytokines and death from cancer. Here, we investigated the association between circulating levels and innate production capacity of proinflammatory cytokines and cancer incidence and mortality in the PROspective Study on Pravastatin in the Elderly at Risk (PROSPER). EXPERIMENTAL DESIGN: Circulating levels of interleukin 6 (IL-6) and C-reactive protein were measured in all 5,804 participants of the PROSPER study. The innate production capacity of IL-6, IL-1beta, and tumor necrosis factor alpha (TNF-alpha) were measured in a random sample of 403 subjects. RESULTS: We showed that high circulating inflammatory markers were associated with an increased risk for cancer incidence and death from cancer during follow-up (all P < 0.05). Moreover, high innate proinflammatory cytokine production capacity is associated with an increased risk for death from cancer (all P < 0.04) but not with higher cancer incidence during follow-up (all P > 0.6). CONCLUSIONS: High innate production capacity of proinflammatory cytokines is associated with an increased risk for death from cancer, probably because of increased tumor growth and metastasis. Because there was no association between innate production capacity and cancer incidence, the association between circulating levels and cancer incidence at least partially reflects reversed causality. (Clin Cancer Res 2009;15(24):7744-8).
Head Neck. 2009 Apr;31(4):429-41.
Lefebvre JL, Ang KK; Larynx Preservation Consensus Panel.
Ang K, Bardet E, Barry B, Bernier J, Bourhis J, Budach V, Calais G, Conley B, Forastiere A, Knecht R, Langendijk J, Lefebvre JL, Leemans R, Licitra L, Posner M, de Raucourt D, Rolland F, Temam S, Vermorken JB, Vokes E, Weber RS, Wolf G.
Département de Cancérologie Cervico-Faciale, Centre Oscar Lambret, Lille, France. jl-lefebvre@o-lambret.fr
BACKGROUND: To develop guidelines for the conduct of phase III clinical trials of larynx preservation in patients with locally advanced laryngeal and hypopharyngeal cancer. METHODS: A multidisciplinary international consensus panel developed recommendations after reviewing results from completed phase III randomized trials, meta-analyses, and published clinical reports with updates available through November 2007. The guidelines were reviewed and approved by the panel. RESULTS: The trial population should include patients with T2 or T3 laryngeal or hypopharyngeal squamous cell carcinoma not considered for partial laryngectomy and exclude those with laryngeal dysfunction or age more than 70 years. Functional assessments should include speech and swallowing. Voice should be routinely assessed with a simple, validated instrument. The primary endpoint should capture survival and function. The panel created a new endpoint: laryngo-esophageal dysfunction-free survival. Events are death, local relapse, total or partial laryngectomy, tracheotomy at 2 years or later, or feeding tube at 2 years or later. Recommended secondary endpoints are overall survival, progression-free survival, locoregional control, time to tracheotomy, time to laryngectomy, time to discontinuation of feeding tube, and quality of life/patient reported outcomes. Correlative biomarker studies for near-term trials should include EGFR, ERCC-1, E-cadherin and beta-catenin, epiregulin and amphiregulin, and TP53 mutation. CONCLUSIONS: Revised trial designs in several key areas are needed to advance the study of larynx preservation. With consistent methodologies, clinical trials can more effectively evaluate and quantify the therapeutic benefit of novel treatment options for patients with locally-advanced laryngeal and hypopharyngeal cancer.
Mol Cancer Ther. 2009 Dec;8(12):3341-9.
Carboni JM, Wittman M, Yang Z, Lee F, Greer A, Hurlburt W, Hillerman S, Cao C, Cantor GH, Dell-John J, Chen C, Discenza L, Menard K, Li A, Trainor G, Vyas D, Kramer R, Attar RM, Gottardis MM.
Oncology Drug Discovery, Department of Pharmaceutical Candidate Organization, Bristol-Myers Squibb Company, PO Box 5400, Princeton, NJ 08543, USA. joan.carboni@bms.com
BMS-754807 is a potent and reversible inhibitor of the insulin-like growth factor 1 receptor/insulin receptor family kinases (Ki, <2 nmol/L). It is currently in phase I development for the treatment of a variety of human cancers. BMS-754807 effectively inhibits the growth of a broad range of human tumor types in vitro, including mesenchymal (Ewing's, rhabdomyosarcoma, neuroblastoma, and liposarcoma), epithelial (breast, lung, pancreatic, colon, gastric), and hematopoietic (multiple myeloma and leukemia) tumor cell lines (IC50, 5-365 nmol/L); the compound caused apoptosis in a human rhabdomyosarcoma cell line, Rh41, as shown by an accumulation of the sub-G1 fraction, as well as by an increase in poly ADP ribose polymerase and Caspase 3 cleavage. BMS-754807 is active in vivo in multiple (epithelial, mesenchymal, and hematopoietic) xenograft tumor models with tumor growth inhibition ranging from 53% to 115% and at a minimum effective dose of as low as 6.25 mg/kg dosed orally daily. Combination studies with BMS-754807 have been done on multiple human tumor cell types and showed in vitro synergies (combination index, <1.0) when combined with cytotoxic, hormonal, and targeted agents. The combination of cetuximab and BMS-754807 in vivo, at multiple dose levels, resulted in improved clinical outcome over single agent treatment. These data show that BMS-754807 is an efficacious, orally active growth factor 1 receptor/insulin receptor family-targeted kinase inhibitor that may act in combination with a wide array of established anticancer agents.
Br J Cancer. 2009 Sep 1;101(5):774-81.
Canner JA, Sobo M, Ball S, Hutzen B, DeAngelis S, Willis W, Studebaker AW, Ding K, Wang S, Yang D, Lin J.
Department of Pediatrics, Nationwide Children's Hospital, Columbus, OH, USA.
BACKGROUND: Interruption of the role of p53s as a tumour suppressor by MDM2 may be one of the mechanisms by which cancer cells evade current therapy. Blocking the inhibition of wild-type p53 by MDM2 in cancer cells should reactivate p53's tumour suppressor functions and enhance current cancer treatments. MI-63 is a novel non-peptide small molecule that has shown strong binding affinity (K(i)=3 nM) for MDM2; however, its effects on paediatric cancer cells and the specific mechanism of tumour suppressor reactivation have not been evaluated. METHODS: Rhabdomyosarcoma (RMS), the most common childhood soft tissue sarcoma, expresses either wild-type or mutant p53 protein. We examined the inhibitory effects of MI-63 in embryonal RMS (ERMS) and alveolar RMS (ARMS) cell lines expressing wild-type or mutated p53. RESULTS: Treatment with MI-63 reduced cell viability by 13.4% and by <1%, respectively, at 72 h in both RH36 and RH18 cell lines expressing wild-type p53. In contrast, RH30 and RD2 cells expressing p53 mutants are resistant to MI-63 treatment.
However, RD2 and RH30 RMS cells, as well as human normal skeletal muscle cells, showed a minimal increase in p53 signalling and no induction of cleaved PARP and caspase-3. MI-63 was compared with Nutlin-3, a known MDM2 inhibitor, and was found to be more potent in the inhibition of cell proliferation/viability. Further, synergy was observed when MI-63 was used in combination with doxorubicin. CONCLUSION: These results indicate that MI-63 is a potent therapeutic agent for RMS cells expressing wild-type p53 protein.
Clin Cancer Res. 2009 Jun 15;15(12):4077-84. Epub 2009 Jun 9.
Miyachi M, Kakazu N, Yagyu S, Katsumi Y, Tsubai-Shimizu S, Kikuchi K, Tsuchiya K, Iehara T, Hosoi H.
Department of Pediatrics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan.
PURPOSE: Seventy to eighty percent of rhabdomyosarcoma (RMS) tumors retain wild-type p53. The tumor suppressor p53 plays a central role in inducing cell cycle arrest or apoptosis in response to various stresses. p53 protein levels are regulated by MDM2 through ubiquitin-dependent degradation. In this study, we evaluated whether nutlin-3, a recently developed small-molecule antagonist of MDM2, has an effect on p53-dependent cell cycle arrest and apoptosis in cultured human RMS cell lines. EXPERIMENTAL DESIGN: Five RMS cell lines with different p53 statuses and MDM2 expression levels were treated with nutlin-3. Gene expression patterns, cell viability, cell cycle, and apoptosis after nutlin-3 treatment, and antitumor activity of combination treatment with vincristine or actinomycin D were assessed. RESULTS: Significant p53 activation was observed in wild-type p53 cell lines after nutlin-3 treatment. p53 activation led to cell cycle arrest in parallel with increased p21 expression. Furthermore, these cell lines underwent p53-dependent apoptosis, concomitant with elevation of proapoptotic genes and activation of caspase-3. The effect of nutlin-3 was almost the same in terms of half maximal inhibitory concentration and apoptosis whether or not MDM2 was overexpressed. Nutlin-3 did not induce either cell cycle arrest or apoptosis in p53 mutant cell lines. A combination of vincristine or actinomycin D with nutlin-3 enhanced the antitumor activity in RMS cell lines with wild-type p53. CONCLUSIONS: Nutlin-3 effectively restored p53 function in both normal MDM2 expression and MDM2 overexpression RMS cell lines with wild-type p53. p53 restoration therapy is a potential therapeutic strategy for refractory RMS with wild-type p53.
Cancer. 2007 Nov 15;110(10):2293-303.
Armistead PM, Salganick J, Roh JS, Steinert DM, Patel S, Munsell M, El-Naggar AK, Benjamin RS, Zhang W, Trent JC.
Department of Sarcoma Medical Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
BACKGROUND: Rhabdomyosarcoma (RMS) is a rare mesenchymal tumor with few treatment options after the failure of first-line therapy. Understanding the expression of kinases and apoptotic molecules in RMS tumors may lead to elucidation of mechanisms of resistance to chemotherapy and development of new therapies. METHODS: Paraffin-embedded tissue samples were collected from 105 RMS patients treated at the M. D. Anderson Cancer Center and examined for the immunohistochemical expression of kinases and apoptotic molecules deemed potential therapeutic targets. Clinicopathologic information was collected on all patients and analyzed for correlation with overall survival (OS). RESULTS: Of the 105 patients, 44 (42%) were female and 89 (85%) were older than 10 years of age. The 5-year OS for this cohort was 24.7%, with inferior median OS in patients with genitourinary primary tumors and those with invasion through the deep fascial plane. Immunohistochemistry revealed Kit and c-erb-b2 to be present on < 10% of tumors but EGFR, PDGFR-alpha, PDGFR-beta, Bcl-2, and Bax were present in > 40% of tumors. Patients whose tumors expressed PDGFR-alpha were found to have a shorter median OS by multivariate analysis (26 vs 266 months, P = .076). Conversely, patients whose tumors expressed Bax were found to have a longer OS (31 vs 19 months, P = .047). CONCLUSIONS: EGFR, PDGFR-alpha, PDGFR-beta, Bcl-2, and Bax are frequently expressed in human RMS tissue and may represent new therapeutic targets. Absence of PDGFR-alpha and the presence of Bax are associated with a longer median OS in patients with RMS. Targeting these molecules may be a successful therapeutic strategy.
Anticancer Res. 2004 Jan-Feb;24(1):249-57.
Staibano S, Franco R, Tranfa F, Mezza E, Lo Muzio L, Strianese D, Errico ME, Bufo P, Ferrara G, Somma P, Mansueto G, Greco I, Fiorillo A, Bonavolontà G, De Rosa G.
Department of Biomorphological and Functional Sciences, Pathology Section, University of Naples Federico II, Italy. amando.tedeschi@virgilio.it
BACKGROUND: As for rhabdomyosarcoma (RMS) of other anatomic regions, the evaluation of traditional clinicopathological parameters does not allow the unequivocal outcome prediction of the single cases of orbital RMS. We investigated the role of DNA ploidy and immunohistochemical expression of p53, bcl-2, MDR-1 and Ki67 (MIB1) in the prognostic evaluation of orbital rhabdomyosarcomas. MATERIALS AND METHODS: The study population consisted of 11 selected cases. Serial sections of each tumor, stained with Feulgen's technique, were analyzed for the DNA content, using the QUANTIMET 500c Leica analyzer, QWINVO200A software. The results were compared with the immunohistochemical expression of p53 (wild plus mutated, W&M and mutated), bcl2, MDR-1 and Ki67 (MIB1), and with follow-up data. RESULTS: The statistical analysis of results showed that the cases of tetraploid and/or multiploid RMS, overexpressing p53 (W&M and mutated) and MDR-1, were characterized by an overall worse prognosis. On the contrary, the tumors with a favourable clinical course showed hyperexpression of MIB1 and absence of mutated p53 expression. Significantly higher MIB1 expression was found in the relapse-free group of tumors, with respect to the RMS with relapse (both in primary tumors and relative relapses, p<0.05). This finding could justify the higher sensibility to pharmacological therapy of RMS of the first group. The group of RMS with a worse prognosis (primary tumors and relapses) showed instead p53 overexpression (W&M and mutated), MDR-1 expression and multiploidy, with high 5cEE values and tetraploid peaks. No significant difference was found concerning the expression of bcl-2 among the two groups of RMS (p>0.05). CONCLUSION: The evaluation of DNA ploidy, p53, MIB1 and MDR-1 expression could be used for subtyping of orbital RMS into two prognostically different subcategories, respectively RMS responder to the therapy, with favourable clinical outcome, and RMS with a worse prognosis, requiring more aggressive therapeutic protocols.
Int J Exp Pathol. 2006 Dec;87(6):445-50.
Department of Veterinary Pathology, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan. imakoto@yamaguchi-u.ac.jp
3-Methylcholanthrene (MC)-induced 10 embryonal (ERSs) and 24 pleomorphic rhabdomyosarcomas (PRSs) of the dermis in mouse were examined immunohistochemically for myogenin, p21 and proliferating cell nuclear antigen (PCNA) nuclear reactivity and myosin reactivity. ERSs had higher expression of myogenin and p21 compared with that of myosin. PRSs were divided into two groups having high (moderate or marked reactivity; HLM) and low (mild reactivity; LLM) levels of myosin expression. Expression of p21 was higher in HLM-PRSs than in LLM-PRSs. Statistically significant association was observed between myosin and p21 expression in PRSs, but not between myosin and myogenin expression. Myogenin and p21 reactivity were observed in myoblast-like cells, but rarely in multinucleated cells. In ERSs, small undifferentiated myogenic precursor cells were also positive for p21. No difference of PCNA reactivity was observed between HLM-PRSs and LLM-PRSs, although its reactivity was higher in PRSs than in ERSs. The results suggest that myogenin is related to myoblast-like cell differentiation in PRSs and that p21 plays essential roles in myotube formation and myosin expression. In ERSs, p21 may be involved in inhibition of myogenic precursor cell proliferation and differentiation.
Nat Immunol. 2010 Apr;11(4):344-9. Epub 2010 Mar 7.
von Essen MR, Kongsbak M, Schjerling P, Olgaard K, Odum N, Geisler C.
Department of International Health, Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark.
Phospholipase C (PLC) isozymes are key signaling proteins downstream of many extracellular stimuli. Here we show that naive human T cells had very low expression of PLC-gamma1 and that this correlated with low T cell antigen receptor (TCR) responsiveness in naive T cells. However, TCR triggering led to an upregulation of approximately 75-fold in PLC-gamma1 expression, which correlated with greater TCR responsiveness. Induction of PLC-gamma1 was dependent on vitamin D and expression of the vitamin D receptor (VDR). Naive T cells did not express VDR, but VDR expression was induced by TCR signaling via the alternative mitogen-activated protein kinase p38 pathway. Thus, initial TCR signaling via p38 leads to successive induction of VDR and PLC-gamma1, which are required for subsequent classical TCR signaling and T cell activation.
Kidney Int. 2010 Feb 24. [Epub ahead of print]
Verstuyf A, Carmeliet G, Bouillon R, Mathieu C.
Laboratory of Experimental Medicine and Endocrinology, Katholieke Universiteit Leuven, Leuven, Belgium.
The secosteroid hormone 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is the natural ligand for the vitamin D receptor, a member of the nuclear receptor superfamily. Upon binding of the ligand, the vitamin D receptor heterodimerizes with the retinoid X receptor and binds to vitamin D response elements in the promoter region of target genes to induce/repress their expression. The target genes that have been identified so far are heterogeneous in nature and reflect the great spectrum of biological activities of 1,25(OH)(2)D(3). Within the last two decades, the receptor has been shown to be present not only in classical target tissues such as bone, kidney, and intestine, but also in many other nonclassical tissues, for example, in the immune system (T and B cells, macrophages, and monocytes), in the reproductive system (uterus, testis, ovary, prostate, placenta, and mammary glands), in the endocrine system (pancreas, pituitary, thyroid, and adrenal cortex), in muscles (skeletal, smooth, and heart muscles), and in brain, skin, and liver. Besides the almost universal presence of vitamin D receptors, different cell types (for example, keratinocytes, monocytes, bone, placenta) are capable of metabolizing 25-hydroxyvitamin D(3) to 1,25(OH)(2)D(3) by the enzyme 25(OH)D(3)-1alpha-hydroxylase, encoded by CYP27B1. The combined presence of CYP27B1 and the specific receptor in several tissues introduced the idea of a paracrine/autocrine role for 1,25(OH)(2)D(3). Moreover, it has been demonstrated that 1,25(OH)(2)D(3) can induce differentiation and inhibit proliferation of normal and malignant cells. Moreover, vitamin D deficiency is associated with an increased risk for nearly all major human diseases such as cancer, autoimmune diseases, cardiovascular, and metabolic diseases. In addition to the treatment of bone disorders with 1,25(OH)(2)D(3), these newly discovered functions open perspectives for the use of 1,25(OH)(2)D(3) as an immune modulator (for example, for the treatment of autoimmune diseases or prevention of graft rejection), inhibitor of cell proliferation, and inducer of cell differentiation (cancer).
Clin Transl Oncol. 2007 Jul;9(7):415-9.
Gallego Melcón S, Sánchez de Toledo Codina J.
Servicio de Oncología y Hematología Pediátrica, Hospital Universitari Vall d'Hebron, Barcelona, Spain. sgallego@vhebron.net
Rhabdomyosarcoma (RMS) is one of the most common extracranial solid tumours in children. Embryonal and alveolar subtypes of RMS present completely different genetic abnormalities. Embryonal RMS (eRMS) is characterised by loss of heterozygosity on the short arm of chromosome 11 (11p15.5), suggesting inactivation of a tumour-suppressor gene. In contrast, the majority (80-85%) of the alveolar RMS (aRMS) have the reciprocal chromosomal translocations 't(2;13)(q35;q14) or t(1;13)(p36;q14). t(2;13) appears in approximately 70% of patients with the alveolar subtype. The molecular counterpart of this translocation consists of the generation of a chimeric fusion gene involving the /PAX3/ gene located in chromosome 2 and a member of the fork-head family, /FOXO1/ (formerly /FKHR/), located in chromosome 13. A less frequent variant translocation t(1;13) involves another PAX family gene, /PAX7/, located in chromosome 1 and /FOXO1/ and is present in 10-15% of cases of the alveolar subtype in RMS. Recently, many studies focused on cancer have demonstrated the great potential of the genomic approach based on tumour expression profiles. These technologies permit the identification of new regulatory pathways. Molecular detection of minimal disease by a sensitive method could contribute to better treatment stratification in these patients. In RMS, the advances in the knowledge of the biological characteristics of the tumour are slowly translated into the clinical management of children with this tumour.
Arch Otolaryngol Head Neck Surg. 2008 May;134(5):522-7.
Lin SK, Shun CT, Kok SH, Wang CC, Hsiao TY, Liu CM.
Department of Dentistry, National Taiwan University Hospital, 1 Chang-Teh St, Taipei, Taiwan.
OBJECTIVE: To verify the inhibitory effects of epigallocatechin-3-gallate (EGCG) on the synthesis of hypoxia-induced vascular endothelial growth factor (VEGF) in nasal polyp fibroblasts (NPFs). DESIGN: Eight primary cultures of NPFs were established from nasal polyps. Effects of EGCG on the production of hypoxia-inducible factor (HIF)-1 alpha (the most potent VEGF stimulant) and VEGF by NPFs under hypoxic conditions were measured by Western blot analysis. Immunohistochemical staining was used to examine the in vivo expressions of HIF-1 alpha and VEGF in 20 sections of nasal polyps. RESULTS: Western blot analysis showed that cobalt chloride induced HIF-1 alpha and VEGF synthesis in NPFs in a time-dependent manner, reaching a plateau at 4 and 8 hours, respectively, following treatment. Epigallocatechin-3-gallate attenuated the level of HIF-1 alpha induced by cobalt chloride and also reduced cobalt chloride-stimulated VEGF production by suppressing HIF-1 alpha synthesis. Furthermore, oligomycin (a specific HIF-1 alpha inhibitor) combined with EGCG resulted in a more profound inhibition of VEGF synthesis compared with oligomycin or EGCG treatment alone. Nevertheless, the synergistic effect seemed smaller than the sum of their individual actions. Immunohistochemical analysis revealed the presence of HIF-1 alpha and VEGF in NPFs and mononuclear round cells. Intimate alignment of VEGF-positive fibroblasts and proliferating small capillaries was frequently found. CONCLUSIONS: Nasal polyp fibroblasts contribute to the pathogenesis of nasal polyps by producing VEGF to promote angiogenesis under hypoxic conditions. Epigallocatechin-3-gallate substantially diminishes HIF-1 alpha and VEGF synthesis in NPFs.
Int J Cancer. 2006 Aug 15;119(4):757-64.
Beevers CS, Li F, Liu L, Huang S.
Department of Biochemistry and Molecular Biology and Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
Curcumin (diferuloylmethane), a polyphenol natural product of the plant Curcuma longa, is undergoing early clinical trials as a novel anticancer agent. However, the anticancer mechanism of curcumin remains to be elucidated. Here we show that curcumin inhibited growth of rhabdomyosarcoma cells (Rh1 and Rh30) (IC50 = 2-5 microM) and arrested cells in G1 phase of the cell cycle. Curcumin also induced apoptosis and inhibited the basal or type I insulin-like growth factor-induced motility of the cells. At physiological concentrations (2.5 microM), curcumin rapidly inhibited phosphorylation of the mammalian target of rapamycin (mTOR) and its downstream effector molecules, p70 S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E (eIF4E) binding protein 1 (4E-BP1), in a panel of cell lines (Rh1, Rh30, DU145, MCF-7 and Hela). Curcumin also inhibited phosphorylation of Akt in the cells, but only at high concentrations (>40 microM). The data suggest that curcumin may execute its anticancer activity primarily by blocking mTOR-mediated signaling pathways in the tumor cells.
Eur J Cancer Prev. 2005 Aug;14(4):351-6.
Chow AW, Murillo G, Yu C, van Breemen RB, Boddie AW, Pezzuto JM, Das Gupta TK, Mehta RG.
Department of Surgical Oncology, College of Medicine, College of Pharmacy, University of Illinois at Chicago, 840 South Wood Street (M/C 820), Chicago, IL 60612, USA.
Rhabdomysarcoma is the most common soft tissue tumour in children under the age of 15. Although the introduction of multimodal treatment programmes, including chemotherapy, radiation therapy and excision have increased the overall survival, the chemotherapeutic agents currently used for the treatment of rhabdomyosarcoma exhibit considerable toxicity. The aim of this study was to investigate the effects and possible mechanism(s) of action of resveratrol on human embryonal rhabdomyosarcoma (RD) cells. Resveratrol is a natural polyphenolic compound produced in a number of edible plants and has received considerable attention as a potential chemopreventive and/or chemotherapeutic agent against various types of cancers. In the present study, resveratrol was shown to inhibit cell proliferation of RD cells in a dose-dependent manner with an IC50 of 48.1 micromol/l and induce an arrest in the S/G2 phase of the cell cycle. As evident from immunocytochemical data, resveratrol treatment increased the size of the RD cells. Furthermore, resveratrol treatment resulted in a significant downregulation of cyclin B expression as demonstrated by western blot analyses. In conclusion, the present study shows that resveratrol exerts a strong inhibition of rhabdomyosarcoma cell proliferation in part by arresting cells in S/G2 phase of the cell cycle. These findings warrant further investigation to establish potential use of resveratrol as a relatively non-toxic chemotherapeutic agent for the treatment of rhabdomyosarcoma.
J Cell Biochem. 2008 Aug 15;104(6):2131-42.
Lin HY, Sun M, Tang HY, Simone TM, Wu YH, Grandis JR, Cao HJ, Davis PJ, Davis FB.
Signal Transduction Laboratory, Ordway Research Institute, 150 New Scotland Avenue, Albany, New York 12208, USA. hlin@ordwayresearch.org
Cyclooxygenase-2 (COX-2) content is increased in many types of tumor cells. We have investigated the mechanism by which resveratrol, a stilbene that is pro-apoptotic in many tumor cell lines, causes apoptosis in human head and neck squamous cell carcinoma UMSCC-22B cells by a mechanism involving cellular COX-2. UMSCC-22B cells treated with resveratrol for 24 h, with or without selected inhibitors, were examined: (1) for the presence of nuclear activated ERK1/2, p53 and COX-2, (2) for evidence of apoptosis, and (3) by chromatin immunoprecipitation to demonstrate p53 binding to the p21 promoter. Stilbene-induced apoptosis was concentration-dependent, and associated with ERK1/2 activation, serine-15 p53 phosphorylation and nuclear accumulation of these proteins. These effects were blocked by inhibition of either ERK1/2 or p53 activation. Resveratrol also caused p53 binding to the p21 promoter and increased abundance of COX-2 protein in UMSCC-22B cell nuclei. Resveratrol-induced nuclear COX-2 accumulation was dependent upon ERK1/2 activation, but not p53 activation. Activation of p53 and p53-dependent apoptosis were blocked by the COX-2 inhibitor, NS398, and by transfection of cells with COX-2-siRNA. In UMSCC-22B cells, resveratrol-induced apoptosis and induction of nuclear COX-2 accumulation share dependence on the ERK1/2 signal transduction pathway. Resveratrol-inducible nuclear accumulation of COX-2 is essential for p53 activation and p53-dependent apoptosis in these cancer cells.
Cancer Res. 1999 Feb 15;59(4):886-94.
Hosoi H, Dilling MB, Shikata T, Liu LN, Shu L, Ashmun RA, Germain GS, Abraham RT, Houghton PJ.
Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105-2794, USA.
The mammalian target of rapamycin (mTOR) has been shown to link growth factor signaling and posttranscriptional control of translation of proteins that are frequently involved in cell cycle progression. However, the role of this pathway in cell survival has not been demonstrated. Here, we report that rapamycin, a specific inhibitor of mTOR kinase, induces G1 cell cycle arrest and apoptosis in two rhabdomyosarcoma cell lines (Rh1 and Rh30) under conditions of autocrine cell growth. To examine the kinetics of rapamycin action, we next determined the rapamycin sensitivity of rhabdomyosarcoma cells exposed briefly (1 h) or continuously (6 days). Results demonstrate that Rh1 and Rh30 cells were equally sensitive to rapamycin-induced growth arrest and apoptosis under either condition. Apoptosis was detected between 24 and 144 h of exposure to rapamycin. Both cell lines have mutant p53; hence, rapamycin-induced apoptosis appears to be a p53-independent process. To determine whether induction of apoptosis by rapamycin was specifically due to inhibition of mTOR signaling, we engineered Rh1 and Rh30 clones to stably express a mutant form of mTOR that was resistant to rapamycin (Ser2035-->Ile; designated mTOR-rr). Rh1 and Rh30 mTOR-rr clones were highly resistant (>3000-fold) to both growth inhibition and apoptosis induced by rapamycin. These results are the first to indicate that rapamycin-induced apoptosis is mediated by inhibition of mTOR. Exogenous insulin-like growth factor (IGF)-I protected both Rh1 and Rh30 from apoptosis, without reactivating ribosomal p70 S6 kinase (p70S6K) downstream of mTOR. However, in rapamycin-treated cultures, the response to IGF-I differed between the cell lines: Rh1 cells proliferated normally, whereas Rh30 cells remained arrested in G1 phase but viable. Rapamycin is known to inhibit synthesis of specific proteins but did not inhibit synthesis or alter the levels of mTOR. To examine the rate at which the mTOR pathway recovered, the ability of IGF-I to stimulate p70S6K activity was followed in cells treated for 1 h with rapamycin and then allowed to recover in medium containing > or =100-fold excess of FK506 (to prevent rapamycin from rebinding to its cytosolic receptor FKBP-12). Our results indicate that, in Rh1 cells, rapamycin dissociates relatively slowly from FKBP-12, with a t1/2 of approximately 17.5 h. in the presence of FK506, whereas there was no recovery of p70S6K activity in the absence of this competitor. This was of interest because rapamycin was relatively unstable under conditions of cell culture having a biological t1/2 of approximately 9.9 h. These results help to explain why cells are sensitive following short exposures to rapamycin and may be useful in guiding the use of rapamycin analogues that are entering clinical trials as novel antitumor agents.
Oncogene. 2004 Nov 18;23(54):8785-95.
Kappler R, Bauer R, Calzada-Wack J, Rosemann M, Hemmerlein B, Hahn H.
Institute of Human Genetics, University of Göttingen, Heinrich-Düker-Weg 12, Göttingen, Germany.
Rhabdomyosarcoma (RMS) is a highly malignant tumor that is histologically related to skeletal muscle, yet genetic and molecular lesions underlying its genesis and progression remain largely unknown. In this study we have compared the molecular profiles of two different mouse models of RMS, each associated with a defined primary genetic defect known to play a role in rhabdomyosarcomagenesis in man. We report that RMS of heterozygous Patched1 (Ptch1) mice show less aggressive growth and a greater degree of differentiation than RMS of heterozygous p53 mice. By means of cDNA microarray analysis we demonstrate that RMS in Ptch1 mutants predominantly express a number of myogenic markers, including myogenic differentiation 1, myosin heavy chain, actin, troponin and tropomyosin, as well as genes associated with Hedgehog/Patched signaling like insulin-like growth factor 2, forkhead box gene Foxf1 and the growth arrest and DNA-damage-inducible gene Gadd45a. In sharp contrast, RMS in p53 mutants display higher expression levels of cell cycle-associated genes like cyclin B1, cyclin-dependent kinase 4 and the proliferation marker Ki-67. These results demonstrate that different causative mutations lead to distinct gene expression profiles in RMS, which appear to reflect their different biological characteristics. Our results provide a first step towards a molecular classification of different forms of RMS. If the described differences can be confirmed in human RMS our results will contribute to a new molecular taxonomy of this cancer, which will be critical for gene mutation- and expression-specific therapy.
Cancer Sci. 2010 Mar 7. [Epub ahead of print]
Wang S, Guo L, Dong L, Guo L, Li S, Zhang J, Sun M.
Department of Pathology, Soochow University School of Medicine, Suzhou, China.
Overexpression of transforming growth factor-beta1 (TGF-beta1) and its downstream molecules in the rhabdomyosarcoma (RMS) RD cell line has been reported previously, but the regulatory role of TGF-beta1 on RMS has not been studied extensively. In the present study, we showed that expression of TGF-beta1 and its downstream molecules type II TGF-beta receptor (TbetaRII) and Smad4 was significantly higher in RMS than in normal skeletal muscle, and there was a significant relationship between TGF-beta1 expression and histological grade. Gene silencing with TGF-beta1 short-hairpin RNA (shRNA)-expressing vectors significantly decreased the growth of RD cells, which was confirmed by caspase-3 (in vitro) and TUNEL (in vivo) assays. Moreover, a proportion of treated rhabdomyosarcoma (RD) cells changed to a round shape from the normal fusiform or polygonal shape and expressed myofilaments. Myogenin is one of the myogenic differentiation genes (MyoD) family of myogenic regulators, and was obviously higher in TGF-beta1-shRNA-treated tumors than it in control at the mRNA and protein level. Immunohistochemical staining with myogenic differentiation markers such as myosin and desmin in subcutaneous RMS tissue showed that TGF-beta1 shRNA increased staining for myosin. These results provide new insight into the biological function of TGF-beta1 in malignant tumors, and imply that the TGF-beta1 signal pathway is a potential therapeutic target for drugs that induce differentiation of RMS. (Cancer Sci 2010).
J Cell Biochem. 2010 May;110(1):162-70.
Gao X, Pan WS, Dai H, Zhang Y, Wu NH, Shen YF.
National Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
CARM1/PRMT4 is a member of the protein arginine methyltransferase (PRMT) family. CARM1 as a transcriptional coactivator plays an active role on mammalian genes. Here, we show that CARM1 can be recruited to the promoter of myogenin gene to enhance its transcriptional activation via PCAF at the early stage of TPA-induced RD cell differentiation. By adding adenosine dialdehyde, AdOx, to inhibit the PRMT in RD cells, the TPA-induced recruiting of p300, PCAF and the Brg1 at the myogenin promoter is abolished and myogenic differentiation is blocked. More specifically, the expression of PCAF and its nucleation are prohibited when CARM1 is knockdown by its specific siRNA. We suggest that the physical interaction of CARM1 and PCAF is likely pivotal for the activation of PCAF in the downstream of CARM1 pathway for inducing myogenin under TPA-induced differentiation. The findings shed lights on novel therapeutic targets in the treatment of rhabdomyosarcoma patients. (c) 2010 Wiley-Liss, Inc.
Am J Surg Pathol. 2001 Sep;25(9):1150-7.
Cessna MH, Zhou H, Perkins SL, Tripp SR, Layfield L, Daines C, Coffin CM.
Department of Pathology, University of Utah School of Medicine, Salt Lake City, USA.
Rhabdomyosarcoma (RMS), the most common soft tissue sarcoma of childhood, displays a variety of histologic patterns. Immunohistochemistry is used extensively to distinguish RMS from its mimics. Myogenin and MyoD1, myogenic transcriptional regulatory proteins expressed early in skeletal muscle differentiation, are considered sensitive and specific markers for RMS and are more specific than desmin and muscle-specific actin and more sensitive than myoglobin. Previous studies have focused on expression of myogenin and MyoD1 in small round cell tumors. This study assesses myogenin and MyoD1 in rhabdomyosarcoma subtypes and spindle cell tumors considered in the differential diagnosis of RMS. Formalin-fixed, paraffin-embedded archival tissue from 32 RMS, 107 non-RMS, and 11 benign skeletal muscle samples was stained for myogenin and MyoD1 with standard immunohistochemical techniques. Nuclear positivity was scored on a three-tiered scale. All RMSs expressed myogenin. Alveolar RMS (ARMS) showed strong nuclear staining, especially in tumor cells lining fibrous septae and perivascular regions. In cases with a subtle alveolar architecture on routinely stained sections, myogenin highlighted and enhanced visualization of the alveolar morphologic pattern. Embryonal RMSs (ERMSs) were more variable in myogenin staining pattern and intensity. No cases of nodular fasciitis, malignant fibrous histiocytoma, malignant peripheral nerve sheath tumor, inflammatory myofibroblastic tumor, myofibrosarcoma, leiomyoma, leiomyosarcoma, or alveolar soft part sarcoma stained for myogenin. Focal nuclear reactivity was seen in desmoid (2 of 10), infantile myofibromatosis (2 of 10), synovial sarcoma (1 of 10), and infantile fibrosarcoma (2 of 10). Non-neoplastic skeletal muscle fiber nuclei stained positively for myogenin in both tumor-associated samples (25 of 40) and benign skeletal muscle samples (5 of 11). Although all RMSs were immunoreactive for MyoD1, cytoplasmic and nonspecific background staining and reactivity of nonmyoid tissues hindered its practical utility in paraffin-embedded samples in this study. Although myogenin is a highly sensitive and specific marker for RMS, it is rarely seen in other spindle cell soft tissue tumors. As previously reported, ARMS stained more strongly than ERMS. In contrast to previous studies, rare non-RMS (7 of 107) displayed focal nuclear reactivity, and entrapped atrophic or regenerative skeletal muscle fibers also stained positively. Although these are potential pitfalls in the interpretation of myogenin, careful attention to morphology and other features, to the relative paucity of myogenin-positive nuclei in non-RMS. and to the presence of entrapped muscle fibers should prevent incorrect interpretation. Because the extent of myogenin expression in RMS is much greater than in non-RMS, it is a very useful marker when interpreted in the context of other clinicopathologic data.
PLoS One. 2010 May 11;5(5):e10578.
Liu L, Chen L, Luo Y, Chen W, Zhou H, Xu B, Han X, Shen T, Huang S.
Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, Louisiana, United States of America.
Serine/threonine (Ser/Thr) protein phosphatase 2A (PP2A) has been implicated as a novel component of the mammalian target of rapamycin (mTOR) signaling pathway. Recently we have demonstrated that mTOR regulates cell motility in part through p70 S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E (eIF4E) binding protein 1 (4E-BP1) pathways. Little is known about the role of PP2A in the mTOR-mediated cell motility. Here we show that rapamycin inhibited the basal or insulin-like growth factor 1 (IGF-1)-induced motility of human Ewing sarcoma (Rh1) and rhabdomyosarcoma (Rh30) cells. Treatment of the cells with rapamycin activated PP2A activity, and concurrently inhibited IGF-1 stimulated phosphorylation of Erk1/2. Inhibition of Erk1/2 with PD98059 did not significantly affect the basal mobility of the cells, but dramatically inhibited IGF-1-induced cell motility. Furthermore, inhibition of PP2A with okadaic acid significantly attenuated the inhibitory effect of rapamycin on IGF-1-stimulated phosphorylation of Erk1/2 as well as cell motility. Consistently, expression of dominant negative PP2A conferred resistance to IGF-1-stimulated phosphorylation of Erk1/2 and cell motility. Expression of constitutively active MKK1 also attenuated rapamycin inhibition of IGF-1-stimulated phosphorylation of Erk1/2 and cell motility. The results suggest that rapamycin inhibits cell motility, in part by targeting PP2A-Erk1/2 pathway.
Cancer Chemother Pharmacol. 2009 Dec 20. [Epub ahead of print]
Zucchetti M, Meco D, Di Francesco AM, Servidei T, Patriarca V, Cusano G, D'Incalci M, Forestieri D, Pisano C, Riccardi R.
Department of Oncology, Istituto di Ricerche Farmacologiche "Mario Negri", Via La Masa 19, 20156, Milan, Italy, zucchetti@marionegri.it.
PURPOSE: This study compared the antitumor activity and the pharmacological profile of gimatecan given orally and irinotecan (CPT-11) on pediatric tumor xenografts. EXPERIMENTAL DESIGN: Gimatecan was tested in two neuroblastoma cell lines (SK-N-DZ and SK-N-(BE)2c) and on TE-671 rhabdomyosarcoma cells using two different schedules. We characterized its pharmacokinetic profile in nude mice bearing human SK-N-DZ and TE-671 cell lines. RESULTS: Gimatecan appears to have high plasma disposition. The drug was present in plasma almost completely as the intact lactone form and showed substantial activity in all tumor models. Prolonged daily treatment with low doses of gimatecan produced significant tumor regression in all tumor xenografts. CONCLUSION: The antitumor activity and the promising pharmacological profile indicate gimatecan as an excellent candidate for clinical treatment of pediatric tumors.
References
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48. PubMed, http://www.ncbi.nlm.nih.gov/pubmed/
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