Genistein inhibits p38 map kinase activation, matrix metalloproteinase type 2, and cell invasion in human prostate epithelial cells.

Epidemiologic studies associate consumption of genistein, in the form of dietary soy, with lower rates of metastatic prostate cancer. We have previously shown that genistein inhibits prostate cancer cell detachment in vitro, that it is well tolerated in an older cohort of men with prostate cancer, and that it alters cell signaling in that same cohort. We have also shown that p38 mitogen-activated protein kinase (MAPK) is necessary for transforming growth factor beta (TGF-beta)-mediated increases in prostate cancer adhesion. Although cell invasion is closely linked to metastatic behavior, little is known about how this process is regulated in prostate cancer or what effect, if any, genistein has on associated processes. We now show that genistein inhibits matrix metalloproteinase type 2 (MMP-2) activity in six of seven prostate cell lines tested, blocks MMP-2 induction by TGF-beta, and inhibits cell invasion. Efficacy was seen at low nanomolar concentrations, corresponding to blood concentrations of free genistein attained after dietary consumption. Inhibition of p38 MAPK by either SB203580 or dominant-negative construct blocked induction of MMP-2 and cell invasion by TGF-beta. Genistein exerted similar effects and was found to block activation of p38 MAPK by TGF-beta. This study shows that p38 MAPK is necessary for TGF-beta-mediated induction of MMP-2 and cell invasion in prostate cancer and that genistein blocks activation of p38 MAPK, thereby inhibiting processes closely linked to metastasis, and does so at concentrations associated with dietary consumption. Any potential causal link to epidemiologic findings will require further investigation.

[1]  G. Bemis,et al.  The structural basis for the specificity of pyridinylimidazole inhibitors of p38 MAP kinase. , 1997, Chemistry & biology.

[2]  L. P. Paul,et al.  Inhibition of Epidermal Growth Factor Receptor-associated Tyrosine Kinase Blocks Glioblastoma Invasion of the Brain. , 1997, Neurosurgery.

[3]  R. Nagle,et al.  Characterization of integrin subunits, cellular adhesion and tumorgenicity of four human prostate cell lines , 1993, Journal of Cancer Research and Clinical Oncology.

[4]  S. Dooley,et al.  In situ hybridization analysis of genes coding collagen IV α1 chain, laminin β1 chain, and s‐laminin in prostate tissue and prostate cancer: Increased basement membrane gene expression in high‐grade and metastatic lesions , 1998, The Prostate.

[5]  C. Takimoto,et al.  Phase I pharmacokinetic and pharmacodynamic analysis of unconjugated soy isoflavones administered to individuals with cancer. , 2003, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[6]  W. Heizer,et al.  Clinical characteristics and pharmacokinetics of purified soy isoflavones: single-dose administration to healthy men. , 2002, The American journal of clinical nutrition.

[7]  J. Supko,et al.  High-performance liquid chromatographic assay for genistein in biological fluids. , 1995, Journal of chromatography. B, Biomedical applications.

[8]  E. Kyle,et al.  Prostate cancer cell growth inhibition by tamoxifen is associated with inhibition of protein kinase C and induction of p21waf1/cip1 , 1998, The Prostate.

[9]  T. Stinchcombe,et al.  Clinical Characteristics and Pharmacokinetics of Purified Soy Isoflavones: Multiple-Dose Administration to Men with Prostate Neoplasia , 2004, Nutrition and cancer.

[10]  S. Barsky,et al.  Genistein inhibits both constitutive and EGF-stimulated invasion in ER-negative human breast carcinoma cell lines. , 1998, Anticancer research.

[11]  E. Kyle,et al.  Genistein-stimulated adherence of prostate cancer cells is associated with the binding of focal adhesion kinase to beta-l-integrin , 1996, Clinical & Experimental Metastasis.

[12]  W. Stetler-Stevenson,et al.  Proteases in invasion: matrix metalloproteinases. , 2001, Seminars in cancer biology.

[13]  E. Kyle,et al.  Focal adhesion kinase (FAK) phosphorylation is not required for genistein-induced FAK-β-1-integrin complex formation , 2004, Clinical & Experimental Metastasis.

[14]  E. Kyle,et al.  Genistein-induced apoptosis of prostate cancer cells is preceded by a specific decrease in focal adhesion kinase activity. , 1997, Molecular pharmacology.

[15]  Jerry L. Adams,et al.  A protein kinase involved in the regulation of inflammatory cytokine biosynthesis , 1994, Nature.

[16]  S. Kambhampati,et al.  p38 MAP kinase modulates Smad-dependent changes in human prostate cell adhesion , 2003, Oncogene.

[17]  C. Legrand,et al.  Transforming growth factor-beta 1 increases the adhesion of MDA-MB-231 mammary adenocarcinoma cells to the microvascular subendothelium. , 1999, Cell adhesion and communication.

[18]  L. Tong,et al.  A highly specific inhibitor of human p38 MAP kinase binds in the ATP pocket , 1997, Nature Structural Biology.

[19]  H. Adlercreutz,et al.  Plasma concentrations of phyto-oestrogens in Japanese men , 1993, The Lancet.

[20]  Kohei Miyazono,et al.  TGF-β signalling from cell membrane to nucleus through SMAD proteins , 1997, Nature.

[21]  H. Adlercreutz.  Western diet and Western diseases: some hormonal and biochemical mechanisms and associations. , 1990, Scandinavian journal of clinical and laboratory investigation. Supplementum.

[22]  M. Schell,et al.  Safety and pharmacokinetics of purified soy isoflavones: single-dose administration to postmenopausal women. , 2002, The American journal of clinical nutrition.

[23]  J. Mohler,et al.  In situ hybridization studies of metalloproteinases 2 and 9 and TIMP-1 and TIMP-2 expression in human prostate cancer , 1997, Clinical & Experimental Metastasis.

[24]  H. Ahn,et al.  Loss of expression of transforming growth factor-beta receptors is associated with poor prognosis in prostate cancer patients. , 1998, Clinical cancer research : an official journal of the American Association for Cancer Research.

[25]  John C. Lee,et al.  Identification of Mitogen-activated Protein (MAP) Kinase-activated Protein Kinase-3, a Novel Substrate of CSBP p38 MAP Kinase (*) , 1996, The Journal of Biological Chemistry.

[26]  S. Schwartz,et al.  Incidence of adenocarcinoma of the prostate in Asian immigrants to the United States and their descendants. , 1999, The Journal of urology.

[27]  Jiahuai Han,et al.  Independent human MAP-kinase signal transduction pathways defined by MEK and MKK isoforms , 1995, Science.

[28]  M. Shibuya,et al.  Genistein, a specific inhibitor of tyrosine-specific protein kinases. , 1987, The Journal of biological chemistry.

[29]  K. Miyazono,et al.  Loss of expression of transforming growth factor beta type I and type II receptors correlates with tumor grade in human prostate cancer tissues. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[30]  C. Takimoto,et al.  Basic pharmacokinetics and pharmacodynamic principles. , 2001, Cancer treatment and research.

[31]  L. Platanias.  Map kinase signaling pathways and hematologic malignancies. , 2003, Blood.

[32]  H. Iishi,et al.  Genistein attenuates peritoneal metastasis of azoxymethane‐induced intestinal adenocarcinomas in Wistar rats , 2000 .

[33]  R. Severson,et al.  A prospective study of demographics, diet, and prostate cancer among men of Japanese ancestry in Hawaii. , 1989, Cancer research.

[34]  Chung Lee,et al.  Genetic change in transforming growth factor β (TGF-β) receptor type I gene correlates with insensitivity to TGF-β1 in human prostate cancer cells , 1996 .

[35]  J. Crowell,et al.  Agent identification and preclinical testing. , 2001, Cancer treatment and research.

[36]  T. Thompson,et al.  Novel regulation of type IV collagenase (matrix metalloproteinase-9 and -2) activities by transforming growth factor-beta1 in human prostate cancer cell lines. , 1999, Molecular biology of the cell.

[37]  G. Stel,et al.  Clinical development plan: Genistein , 2002 .

[38]  P. Kostenuik,et al.  Transforming growth factor β upregulates the integrin-mediated adhesion of human prostatic carcinoma cells to type I collagen , 2004, Clinical & Experimental Metastasis.

[39]  Carlos López-Otín,et al.  Strategies for MMP inhibition in cancer: innovations for the post-trial era , 2002, Nature Reviews Cancer.

[40]  C. Yan,et al.  Suppression of adhesion-induced protein tyrosine phosphorylation decreases invasive and metastatic potentials of B16-BL6 melanoma cells by protein tyrosine kinase inhibitor genistein. , 1997, Invasion & metastasis.

[41]  J. Massagué.  TGFβ Signaling: Receptors, Transducers, and Mad Proteins , 1996, Cell.

[42]  T. Thompson,et al.  Transforming growth factor beta1 stimulates contrasting responses in metastatic versus primary mouse prostate cancer-derived cell lines in vitro. , 1996, Cancer research.

[43]  A. Angelucci,et al.  Osteoblast conditioned media contain TGF‐β1 and modulate the migration of prostate tumor cells and their interactions with extracellular matrix components , 1999, International journal of cancer.

[44]  B. Henderson,et al.  Cancers of the prostate and breast among Japanese and white immigrants in Los Angeles County. , 1991, British Journal of Cancer.

[45]  D. Grignon,et al.  Membrane type 1-matrix metalloproteinase (MT1-MMP) and MMP-2 immunolocalization in human prostate: change in cellular localization associated with high-grade prostatic intraepithelial neoplasia. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[46]  C. Yan,et al.  Genistein suppresses adhesion-induced protein tyrosine phosphorylation and invasion of B16-BL6 melanoma cells. , 1998, Cancer letters.

[47]  Chung Lee,et al.  Over expression of endoglin in human prostate cancer suppresses cell detachment, migration and invasion , 2002, Oncogene.

[48]  E. Kyle,et al.  Prostate cancer chemoprevention agents exhibit selective activity against early stage prostate cancer cells , 2001, Prostate Cancer and Prostatic Diseases.

[49]  R. Weinberg,et al.  Biochemical evidence for the autophosphorylation and transphosphorylation of transforming growth factor beta receptor kinases. , 1995, Proceedings of the National Academy of Sciences of the United States of America.