Investigational New Drugs

, Volume 22, Issue 2, pp 107–117 | Cite as

Resveratrol Antagonizes EGFR-Dependent Erk1/2 Activation in Human Androgen-Independent Prostate Cancer Cells with Associated Isozyme-Selective PKCα Inhibition

  • Jubilee R. Stewart
  • Catherine A. O'Brian


The development of androgen-independent prostate cancer (AI PrCa) involves constitutive Erk1/2 activation sustained by the epidermal growth factor/transforming growth factor-α/EGF receptor (EGF/TGFα/EGFR) axis and other trophic signaling mechanisms in neoplastic human prostate epithelial cells in vivo. In this report, we show that growth-inhibitory concentrations of the dietary phytochemical resveratrol suppress EGFR-dependent Erk1/2 activation pathways stimulated by EGF and phorbol ester (12-O-tetradecanoyl phorbol 13-acetate, TPA) in human AI PrCa PC-3 cells in vitro. Because protein kinase C (PKC) is the major cellular receptor for phorbol esters and taking into consideration that resveratrol is PKC-inhibitory, we investigated resveratrol effects on cellular PKC isozymes associated with the suppression of TPA-induced Erk1/2 activation. The PKC isozyme composition of PC-3 cells was defined by Western analysis of the cell lysate with a comprehensive set of isozyme-selective PKC Ab's. PC-3 cells expressed PKCα, ε, ζ, ι, and PKD (PKCμ), as did another human AI PrCa cell line of distinct genetic origin, DU145. The effects of resveratrol on TPA-induced PKC isozyme activation were defined by monitoring PKC isozyme translocation and autophosphorylation. Under conditions where resveratrol suppressed TPA-induced Erk1/2 activation, the phytochemical produced isozyme-selective interference with TPA-induced translocation of cytosolic PKCα to the membrane/cytoskeleton and selectively diminished the amount of autophosphorylated PKCα in the membrane/cytoskeleton of the TPA-treated cells. These results demonstrate that resveratrol abrogation of a PKC-mediated Erk1/2 activation response in PC-3 cells correlates with isozyme-selective PKCα inhibition. The results provide evidence that resveratrol may have value as an adjuvant cancer therapeutic in advanced prostate cancer.

resveratrol protein kinase C-alpha (PKCα) Erk1/2 androgen-independent prostate cancer epidermal growth factor receptor 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Yang CS, Landau JM, Huang M-T, Newmark HL: Inhibition of carcinogenesis by dietary polyphenolic compounds. Annu Rev Nutr 21: 381–406, 2001Google Scholar
  2. 2.
    Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CWW, Fong HHS, Farnsworth NR, Kinghom AD, Mehta RG, Moon RC, Pezzuto JM: Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275: 218–220, 1997Google Scholar
  3. 3.
    Gusman J, Malonne H, Atassi G: A reappraisal of the potential chemopreventive and chemotherapeutic properties of resveratrol. Carcinogenesis 22: 1111–1117, 2001.Google Scholar
  4. 4.
    Subbaramaiah K, Chung WJ, Michaluart P, Telang N, Tanabe T, Inoue H, Jang M, Pezzuto JM, Dannenberg AJ: Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells. JBC 273: 21875–21882, 1998Google Scholar
  5. 5.
    Mutoh M, Takahashi M, Fukuda K, Matsushima-Hibiya Y, Hiroshi Mutoh H, Sugimura T, Wakabayashi K: Suppression of cyclooxygenase-2 promoter-dependent transcriptional activity in colon cancer cells by chemopreventive agents with a resorcin-type structure. Carcinogenesis 21: 959–963, 2000.Google Scholar
  6. 6.
    Sgambato A, Ardito R, Faraglia B, Boninsegna A, Wolf FI, Cittadini A: Resveratrol, a natural phenolic compound, inhibits cell proliferation and prevents oxidative DNA damage. Mutat Res 496: 171–180, 2001Google Scholar
  7. 7.
    Hsieh T-C, Wu JM: Differential effects on growth, cell cycle arrest, and induction of apoptosis by resveratrol in human prostate cancer cell lines. Exp Cell Res 249: 109–115, 1999Google Scholar
  8. 8.
    Schneider Y, Vincent F, Duranton B, Badolo L, Gossé F, Bergmann C, Seiler N, Raul F: Anti-proliferative effect of resveratrol, a natural component of grapes and wine, on human colonic cancer cells. Cancer Lett 158: 85–91, 2000Google Scholar
  9. 9.
    Lara PN, Jr., Meyers FJ: Treatment options in androgen-independent prostate cancer. Cancer Inves 17: 137–144, 1999Google Scholar
  10. 10.
    Scher HI, Sarkis A, Reuter V, Cohen D, Netto G, Petrylak D, Lianes P, Fuks Z, Mendelsohn J, Cordon-Cardo C: Changing pattern of expression of the epidermal growth factor receptor and transforming growth factor alpha in the progression of prostatic neoplasms. Clin Cancer Res 1: 545–550, 1995Google Scholar
  11. 11.
    Jones HE, Eaton CL, Barrow D, Dutkowski CM, Gee JMW, Griffiths K: Comparative studies of the mitogenic effects of epidermal growth factor and transforming growth factor-and the expression of various growth factors in neoplastic and non-neoplastic prostatic cell lines. Prostate 30: 219–231, 1997Google Scholar
  12. 12.
    Prewett M, Rockwell P, Rockwell RF, Giorgio NA, Mendelsohn J, Scher HI, Goldstein NI: The biologic effects of C225, a chimeric monoclonal antibody to the EGFR, on human prostate carcinoma. J Immunother Emphasis Tumor Immunol 19: 419–427, 1996Google Scholar
  13. 13.
    Hofer DR, Sherwood ER, Bromberg WD, Mendelsohn J, Lee C, Kozlowski JM: Autonomous growth of androgen-independent human prostatic carcinoma cells: role of transforming growth factor alpha. Cancer Res 51: 2780–2785, 1991Google Scholar
  14. 14.
    Culig Z, Hobisch A, Cronauer MV, Radmayr C, Trapman J, Hittmair A, Bartsch G, Klocker H: Androgen receptor activation in prostatic tumor cell lines by insulin-like growth factor-I, keratinocyte growth factor, and epidermal growth factor. Cancer Res. 54: 5474–5478, 1994Google Scholar
  15. 15.
    Robinson MJ, Cobb MH: Mitogen-activated protein kinase pathways. Curr Opin Cell Biol 9: 180–186, 1997Google Scholar
  16. 16.
    Putz T, Culig Z, Eder IE, Nessler-Menardi C, Bartsch G, Grunicke H, Überall F, Klocker H: Epidermal growth factor (EGF) receptor blockade inhibits the Action of EGF, insulin-like growth factor I, and a protein kinase A activator on the mitogen-activated protein kinase pathway in prostate cancer cell lines. Cancer Res 59: 227–233, 1999Google Scholar
  17. 17.
    Yeh S, Lin H-K, Kang H-Y, Thin TH, Lin M-F, Chang C: From HER2/Neu signal cascade to androgen receptor and its coactivators: a novel pathway by induction of androgen target genes through MAP kinase in prostate cancer cells. PNAS 96: 5458–5463, 1999Google Scholar
  18. 18.
    Gioeli D, Mandell JW, Petroni GR, Frierson HF, Jr., Web MJ: Activation of mitogen-activated protein kinase associated with prostate cancer progression. Cancer Res 59: 279–284, 1999Google Scholar
  19. 19.
    Price D, Rocca G, Gou C, Ballo M, Schwinn D, Luttrell L: Activation of extracellular signal-regulated kinase in human prostate cancer. J Urol 162: 1537–1542, 1999Google Scholar
  20. 20.
    Chen N, Ma W-Y, She Q-B, Wu E, Liu G, Bode AM, Dong Z: Transactivation of the epidermal growth factor receptor is involved in 12-O-Tetradecanoylphorbol-13-acetate-induced signal transduction. JBC 276: 46722–46728, 2001.Google Scholar
  21. 21.
    Newton AC: Regulation of protein kinase C. Cur Opin Cell Biol 9: 161–167, 1997Google Scholar
  22. 22.
    Stewart JR, Ward NE, Ioannides CG, O'Brian CA: Resveratrol preferentially inhibits protein kinase C-catalyzed phosphorylation of a cofactor-independent, arginine-rich protein substrate by a novel mechanism. Biochemistry 38: 13244–13251, 1999Google Scholar
  23. 23.
    Stewart JR, Christman KL, O'Brian CA: Effects of resveratrol on the autophosphorylation of phorbol ester-responsive protein kinases: inhibition of protein kinase D but not protein kinase C isozyme autophosphorylation. Biochem Pharmacol 60: 1355–1359, 2000Google Scholar
  24. 24.
    Ács P, Bögi K, Marquez AM, Bíró T, Szállási Z, Blumberg PM: The catalytic domain of protein kinase C chimeras modulates the affinity and targeting of phorbol ester-induced translocation. JBC 272: 22148–22153, 1997Google Scholar
  25. 25.
    Ogretmen B, Schady D, Usta J, Wood R, Kraveka JM, Luberto C, Birbes H, Hannun YA, Obeid LM: Role of ceramide in mediating the inhibition of telomerase activity in A549 human lung adenocarcinoma cells. JBC 276: 24901–24910, 2001Google Scholar
  26. 26.
    Gupta KP, Ward NE, Gravitt KR, Bergman PJ, O'Brian CA: Partial reversal of multidrug resistance in human breast cancer cells by an N-myristoylated protein kinase C-α pseudosubstrate peptide. JBC 271: 2102–2111, 1996Google Scholar
  27. 27.
    De Miguel P, Royuela, Bethencourf R, Ruiz, Fraile, Paniagua: Immunohistochemical comparative analysis of transforming growth factor, epidermal growth factor, and epidermal growth factor receptor in normal, hyperplastic and neoplastic human prostates. Cytokine 11: 722–727, 1999Google Scholar
  28. 28.
    Gou C, Luttrell L, Price D: Mitogenic signaling in androgen sensitive and insensitive prostate cancer cell lines. J Urol 163: 1027–1032, 2000Google Scholar
  29. 29.
    Payne DM, Rossomando AJ, Martino P, Erickson AK, Her JH, Shabanowitz J, Hunt DF, Weber MJ, Sturgill TW: Identification of the regulatory phosphorylation sites in pp42/mitogen-activated protein kinase (MAP kinase). EMBO J 10: 885–892, 1991Google Scholar
  30. 30.
    Sturgill TW et al.: Nature 334: 715–718, 1988Google Scholar
  31. 31.
    Kolch W, Heldecker G, Kochs G, Hummel R, Vahidi H, Mischak H, Finkenzeller G, Marme D, Rapp UR: Protein kinase C activates RAF-1 by direct phosphorylation. Nature 364: 249–252, 1993.Google Scholar
  32. 32.
    Miranti CK, Ohno S, Brugge JS: Protein kinase C regulates integrin-induced activation of the extracellular regulated kinase pathway upstream of Shc. JBC 274: 10571–10581, 1999Google Scholar
  33. 33.
    Comford P, Evans J, Dodson A, Parsons K, Woolfenden A, Neoptolemos J, Foster CS: Protein kinase C isoenzyme patterns characteristically modulated in early prostate cancer. Am J Pathol 154: 137–144, 1999Google Scholar
  34. 34.
    Ron D, Kazanietz M: New insights into the regulation of protein kinase C and novel phorbol ester receptors. FASEB J 13: 1658–1676, 1999.Google Scholar
  35. 35.
    Matthews SA, Rozengurt E, Cantrell D: Characterization of serine 916 as an in vivo autophosphorylation site for protein kinase D/protein kinase Cμ. JBC 274: 26543–26549, 1999Google Scholar
  36. 36.
    Vertommen D, Rider M, Ni Y, Waelkens E, Merlevede W, Vandenheede JR, Van Lint J: Regulation of protein kinase D by multisite phosphorylation: identification of phosphorylation sites by mass spectrometry and characterization by site-directed mutagenesis. JBC 275: 19567–19576, 2000Google Scholar
  37. 37.
    Ahmad N, Adhami VM, Afaq F, Feyes DK, Mukhtar H: Resveratrol causes WAF-1/p21-mediated G1-phase arrest of cell cycle and induction of apoptosis in human epidermoid carcinoma A431 Cells. Clin Cancer Res 7: 1466–1473, 2001Google Scholar
  38. 38.
    Mouria M, Gukovskaya AS, Jung Y, Buechler P, Hines OJ, Reber HA, Pandol SJ: Food-derived polyphenols inhibit pancreatic cancer growth through mitochondrial cytochrome C release and apoptosis. Int J Cancer 98: 761–769, 2002.Google Scholar
  39. 39.
    Gehm BD, McAndrews JM, Chien P-Y, Jameson JL: Resveratrol, a polyphenolic compound found in grapes and wine, is an agonist for the estrogen receptor. PNAS 94: 14138–14143, 1997Google Scholar
  40. 40.
    Bhat KPL, Lantvit D, Christov K, Mehta RG, Moon RC, Pezzuto JM: Estrogenic and antiestrogenic properties of resveratrol in mammary tumor models. Cancer Res 61: 7456–7463, 2001Google Scholar
  41. 41.
    Mgbonyebi OP, Russo J, Russo IH: Antiproliferative effect of synthetic resveratrol on human breast epithelial cells. Int J Oncol 12: 865–869, 1998Google Scholar
  42. 42.
    Mitchell SH, Zhu W, Young CYF: Resveratrol inhibits the expression and function of the androgen receptor in LNCaP prostate cancer cells. Cancer Res 59: 5892–5895, 1999Google Scholar
  43. 43.
    Logothetis CJ: Docetaxel in the integrated management of prostate cancer. Current applications and future promise. Oncology 16: 63–72, 2002Google Scholar
  44. 44.
    Karashima T, Sweeney P, Slaton JW, Kim SJ, Kedar D, Izawa JI, Fan Z, Pettaway C, Hicklin DJ, Shuin T, Dinney CP: Inhibition of angiogenesis by the antiepidermal growth factor receptor antibody ImClone C225 in androgen-independent prostate cancer growing orthotopically in nude mice. Clin Cancer Res 8: 1253–1264, 2002.Google Scholar
  45. 45.
    Sirotnak FM, Zakowski MF, Miller VA, Scher HI, Kris MG, Efficacy of cytotoxic agents against human tumor xenografts is markedly enhanced by coadministration of ZD1839 (Iressa), an inhibitor of EGFR tyrosine kinase. Clin Cancer Res 6: 4885–4892, 2000Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Jubilee R. Stewart
    • 1
  • Catherine A. O'Brian
    • 1
  1. 1.Department of Cancer BiologyU.T.M.D. Anderson Cancer CenterHoustonU.S.A

Personalised recommendations