The inhibitory effects of 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone on human colon cancer cells.

SCOPE Previously, we reported that 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF), a polymethoxyflavone found in citrus peels, potently inhibited the growth of multiple human colon cancer cells. Herein, we further investigated the anti-cancer mechanisms of 5HHMF in human colon cancer cells. METHODS AND RESULTS Colony formation assay revealed that 5HHMF dose dependently inhibited colony formation of multiple colon cancer cells. Western blot analysis demonstrated 5HHMF decreased nuclear β-catenin levels and increased the E-cadherin level in a dose-dependent manner. 5HHMF also modified plasma membrane-associated proteins, such as K-Ras, EGFR, and their downstream effectors, such as Akt. Moreover, treatments with 5HHMF inhibited nuclear translocation of NF-κB, which may contribute to its anti-cancer effects. Add-back study showed that the inhibitory effect of 5HHMF was not associated with the production of reactive oxygen species (ROS). In addition, 5HHMF treatment inhibited the capillary tube formation of human umbilical vein endothelial cells (HUVECs) on matrigel, suggesting a potential anti-tumor angiogenesis effect. CONCLUSION Our results demonstrated that 5HHMF suppressed multiple oncogenic molecular events in colon cancer cells.

[1]  D. Mcclements,et al.  The p53-, Bax- and p21-dependent inhibition of colon cancer cell growth by 5-hydroxy polymethoxyflavones. , 2011, Molecular nutrition & food research.

[2]  P. Harari,et al.  Understanding resistance to EGFR inhibitors—impact on future treatment strategies , 2010, Nature Reviews Clinical Oncology.

[3]  D. Mcclements,et al.  Inhibitory effects of 5-hydroxy polymethoxyflavones on colon cancer cells. , 2010, Molecular nutrition & food research.

[4]  A. Papavassiliou,et al.  Colon carcinogenesis: Learning from NF-kappaB and AP-1. , 2010, The international journal of biochemistry & cell biology.

[5]  E. Van Cutsem,et al.  Cetuximab and chemotherapy as initial treatment for metastatic colorectal cancer. , 2009, The New England journal of medicine.

[6]  Qing‐Yu He,et al.  Identification and characterization of molecular targets of natural products by mass spectrometry. , 2009, Mass spectrometry reviews.

[7]  Chi-Tang Ho,et al.  Monodemethylated polymethoxyflavones from sweet orange (Citrus sinensis) peel inhibit growth of human lung cancer cells by apoptosis. , 2009, Molecular nutrition & food research.

[8]  Han-Ming Shen,et al.  NFκB signaling in carcinogenesis and as a potential molecular target for cancer therapy , 2009, Apoptosis.

[9]  Dongsheng Tu,et al.  K-ras mutations and benefit from cetuximab in advanced colorectal cancer. , 2008, The New England journal of medicine.

[10]  Daniel J. Freeman,et al.  Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[11]  E. Van Cutsem,et al.  KRAS wild-type state predicts survival and is associated to early radiological response in metastatic colorectal cancer treated with cetuximab. , 2008, Annals of oncology : official journal of the European Society for Medical Oncology.

[12]  D. Gorski,et al.  Regulation of angiogenesis through a microRNA (miR-130a) that down-regulates antiangiogenic homeobox genes GAX and HOXA5. , 2008, Blood.

[13]  Chi-Tang Ho,et al.  Apoptosis-inducing activity of hydroxylated polymethoxyflavones and polymethoxyflavones from orange peel in human breast cancer cells. , 2007, Molecular nutrition & food research.

[14]  M. Červinka,et al.  Polyphenolic compounds in chemoprevention of colon cancer - targets and signaling pathways. , 2007, Anti-cancer agents in medicinal chemistry.

[15]  Ying-Jan Wang,et al.  5-Hydroxy-3,6,7,8,3',4'-hexamethoxyflavone induces apoptosis through reactive oxygen species production, growth arrest and DNA damage-inducible gene 153 expression, and caspase activation in human leukemia cells. , 2007, Journal of agricultural and food chemistry.

[16]  T. Frebourg,et al.  Clinical relevance of KRAS mutation detection in metastatic colorectal cancer treated by Cetuximab plus chemotherapy , 2007, British Journal of Cancer.

[17]  Sejal Patel,et al.  The Homeobox Gene GAX Activates p21WAF1/CIP1 Expression in Vascular Endothelial Cells through Direct Interaction with Upstream AT-rich Sequences* , 2007, Journal of Biological Chemistry.

[18]  Chi-Tang Ho,et al.  Polymethoxylated flavones induce Ca(2+)-mediated apoptosis in breast cancer cells. , 2006, Life sciences.

[19]  I. Tomlinson,et al.  Colorectal cancer and genetic alterations in the Wnt pathway , 2006, Oncogene.

[20]  Chi-Tang Ho,et al.  Comparison of supercritical fluid chromatography and liquid chromatography for the separation of urinary metabolites of nobiletin with chiral and non-chiral stationary phases. , 2006, Biomedical chromatography : BMC.

[21]  Chi-Tang Ho,et al.  Hydroxylated polymethoxyflavones and methylated flavonoids in sweet orange (Citrus sinensis) peel. , 2006, Journal of agricultural and food chemistry.

[22]  A. Lièvre,et al.  KRAS mutation status is predictive of response to cetuximab therapy in colorectal cancer. , 2006, Cancer research.

[23]  Sejal Patel,et al.  The homeobox gene Gax inhibits angiogenesis through inhibition of nuclear factor-kappaB-dependent endothelial cell gene expression. , 2005, Cancer research.

[24]  T. Venesio,et al.  Near‐diploid and near‐triploid human sporadic colorectal adenocarcinomas differ for KRAS2 and TP53 mutational status , 2003, Genes, chromosomes & cancer.

[25]  D. Gorski,et al.  Inhibition of endothelial cell activation by the homeobox gene Gax. , 2003, The Journal of surgical research.

[26]  Ho-Jin Park,et al.  3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Inhibitors Interfere With Angiogenesis by Inhibiting the Geranylgeranylation of RhoA , 2002, Circulation research.

[27]  J. Coxhead,et al.  Mutations in APC, Kirsten-ras, and p53—alternative genetic pathways to colorectal cancer , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Avri Ben-Ze'ev,et al.  The cadherin-catenin adhesion system in signaling and cancer. , 2002, The Journal of clinical investigation.

[29]  Gerhard Christofori,et al.  Molecular Mechanisms of Tumor Angiogenesis and Tumor Progression , 2000, Journal of Neuro-Oncology.

[30]  A L Zhou,et al.  Matrix metalloproteinase activity is required for activity-induced angiogenesis in rat skeletal muscle. , 2000, American journal of physiology. Heart and circulatory physiology.

[31]  A. Harłozińska,et al.  Expression of the Tyrosine Kinase Activity Growth Factor Receptors (EGFR, ERB B2, ERB B3) in Colorectal Adenocarcinomas and Adenomas , 2000, Tumor Biology.

[32]  O. Sansom,et al.  Dysregulated expression of β-catenin marks early neoplastic change in Apc mutant mice, but not all lesions arising in Msh2 deficient mice , 1999, Oncogene.

[33]  S. Orsulic,et al.  E-cadherin binding prevents beta-catenin nuclear localization and beta-catenin/LEF-1-mediated transactivation. , 1999, Journal of cell science.

[34]  A. Norman,et al.  Kirsten ras mutations in patients with colorectal cancer: the multicenter "RASCAL" study. , 1998, Journal of the National Cancer Institute.

[35]  Gerhard Christofori,et al.  A causal role for E-cadherin in the transition from adenoma to carcinoma , 1998, Nature.

[36]  M. Sporn,et al.  Recent advances in chemoprevention of cancer. , 1997, Science.

[37]  H. Aberle,et al.  Cadherin‐catenin complex: Protein interactions and their implications for cadherin function , 1996, Journal of cellular biochemistry.

[38]  H. Ludwig,et al.  The prognostic significance of proliferating cell nuclear antigen, epidermal growth factor receptor, and mdr gene expression in colorectal cancer , 1993, Cancer.

[39]  C. Rao,et al.  Molecular markers and targets for colorectal cancer prevention , 2008, Acta Pharmacologica Sinica.

[40]  A. Jemal,et al.  Cancer Statistics, 2007 , 2007, CA: a cancer journal for clinicians.

[41]  A. Baldwin,et al.  Nuclear factor-kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. , 2006, Oncogene.

[42]  F. Russo,et al.  EGF, TGF-a, and EGF-R in Human Colorectal Adenocarcinoma , 1998 .

[43]  F. Russo,et al.  EGF, TGF-alpha, and EGF-R in human colorectal adenocarcinoma. , 1998, Acta oncologica.

[44]  J. Folkman.  Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.

[45]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.

[46]  William E. Grizzle,et al.  Detection of high incidence of K-ras oncogenes during human colon tumorigenesis , 1987, Nature.

[47]  B. Vogelstein,et al.  Prevalence of ras gene mutations in human colorectal cancers , 1987, Nature.