Molecular and Cellular Biochemistry

, Volume 304, Issue 1–2, pp 273–285 | Cite as

Molecular mechanisms of resveratrol (3,4,5-trihydroxy-trans-stilbene) and its interaction with TNF-related apoptosis inducing ligand (TRAIL) in androgen-insensitive prostate cancer cells

  • Sharmila Shankar
  • Imtiaz Siddiqui
  • Rakesh K. Srivastava


Although resveratrol, an active ingredient derived from grapes and red wine, possesses chemopreventive properties against several cancers, the molecular mechanisms by which it inhibits cell growth and induces apoptosis have not been clearly understood. Here, we examined the molecular mechanisms of resveratrol and its interactive effects with TRAIL on apoptosis in prostate cancer PC-3 and DU-145 cells. Resveratrol inhibited cell viability and colony formation, and induced apoptosis in prostate cancer cells. Resveratrol downregulated the expression of Bcl-2, Bcl-XL and survivin and upregulated the expression of Bax, Bak, PUMA, Noxa, and Bim, and death receptors (TRAIL-R1/DR4 and TRAIL-R2/DR5). Treatment of prostate cancer cells with resveratrol resulted in generation of reactive oxygen species (ROS), translocation of Bax to mitochondria and subsequent drop in mitochondrial membrane potential, release of mitochondrial proteins (cytochrome c, Smac/DIABLO, and AIF) to cytosol, activation of effector caspase-3 and caspase-9, and induction of apoptosis. Resveratrol-induced ROS production, caspase-3 activity and apoptosis were inhibited by N-acetylcysteine. Bax was a major proapoptotic gene mediating the effects of resveratrol as Bax siRNA inhibited resveratrol-induced apoptosis. Resveratrol enhanced the apoptosis-inducing potential of TRAIL, and these effects were inhibited by either dominant negative FADD or caspase-8 siRNA. The combination of resveratrol and TRAIL enhanced the mitochondrial dysfunctions during apoptosis. These properties of resveratrol strongly suggest that it could be used either alone or in combination with TRAIL for the prevention and/or treatment of prostate cancer.


Bcl-2 Mitochondria IAP TRAIL Cytochrome c Apoptosis Prostate cancer Caspase 



This work was supported by the National Institutes of Health. We thank Dr. Vishva Dixit (Genentech, South San Francisco, CA) for providing dominant negative FADD.


  1. 1.
    Landis SH, Murray T, Bolden S, Wingo PA (1999) Cancer statistics, 1999. CA Cancer J Clin 49:8–31, 1Google Scholar
  2. 2.
    Long RJ, Roberts KP, Wilson MJ, Ercole CJ, Pryor JL (1997) Prostate cancer: a clinical and basic science review. J Androl 18:15–20PubMedGoogle Scholar
  3. 3.
    Petrylak DP (1999) Chemotherapy for advanced hormone refractory prostate cancer. Urology 54:30–35PubMedCrossRefGoogle Scholar
  4. 4.
    Pisters LL (1999) The challenge of locally advanced prostate cancer. Semin Oncol 26:202–216PubMedGoogle Scholar
  5. 5.
    Boyle P, Maisonneuve P, Napalkov P (1995) Geographical and temporal patterns of incidence and mortality from prostate cancer. Urology 46:47–55PubMedCrossRefGoogle Scholar
  6. 6.
    Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CW, Fong HH, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM (1997) Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275:218–220PubMedCrossRefGoogle Scholar
  7. 7.
    Cos P, De Bruyne T, Apers S, Vanden Berghe D, Pieters L, Vlietinck AJ (2003) Phytoestrogens: recent developments. Planta Med 69:589–599PubMedCrossRefGoogle Scholar
  8. 8.
    Inoue H, Jiang XF, Katayama T, Osada S, Umesono K, Namura S (2003) Brain protection by resveratrol and fenofibrate against stroke requires peroxisome proliferator-activated receptor alpha in mice. Neurosci Lett 352:203–206PubMedCrossRefGoogle Scholar
  9. 9.
    Agarwal R (2000) Cell signaling and regulators of cell cycle as molecular targets for prostate cancer prevention by dietary agents. Biochem Pharmacol 60:1051–1059PubMedCrossRefGoogle Scholar
  10. 10.
    Hsieh TC, Wu JM (2000) Grape-derived chemopreventive agent resveratrol decreases prostate-specific antigen (PSA) expression in LNCaP cells by an androgen receptor (AR)-independent mechanism. Anticancer Res 20:225–228PubMedGoogle Scholar
  11. 11.
    Bhat KP, Pezzuto JM (2002) Cancer chemopreventive activity of resveratrol. Ann NY Acad Sci 957:210–229PubMedCrossRefGoogle Scholar
  12. 12.
    Garvin S, Ollinger K, Dabrosin C (2006) Resveratrol induces apoptosis and inhibits angiogenesis in human breast cancer xenografts in vivo. Cancer Lett 231:113–122PubMedCrossRefGoogle Scholar
  13. 13.
    Delmas D, Rebe C, Micheau O, Athias A, Gambert P, Grazide S, Laurent G, Latruffe N, Solary E (2004) Redistribution of CD95, DR4 and DR5 in rafts accounts for the synergistic toxicity of resveratrol and death receptor ligands in colon carcinoma cells. Oncogene 23:8979–8986PubMedCrossRefGoogle Scholar
  14. 14.
    Fulda S, Debatin KM (2004) Sensitization for anticancer drug-induced apoptosis by the chemopreventive agent resveratrol. Oncogene 23:6702–6711PubMedCrossRefGoogle Scholar
  15. 15.
    Fulda S, Debatin KM (2005) Resveratrol-mediated sensitisation to TRAIL-induced apoptosis depends on death receptor and mitochondrial signalling. Eur J Cancer 41:786–798PubMedCrossRefGoogle Scholar
  16. 16.
    Garg AK, Buchholz TA, Aggarwal BB (2005) Chemosensitization and radiosensitization of tumors by plant polyphenols. Antioxid Redox Signal 7:1630–1647PubMedCrossRefGoogle Scholar
  17. 17.
    Srivastava RK (2000) Intracellular mechanisms of TRAIL and its role in cancer therapy. Mol Cell Biol Res Commun 4:67–75PubMedCrossRefGoogle Scholar
  18. 18.
    Suliman A, Lam A, Datta R, Srivastava RK (2001) Intracellular mechanisms of TRAIL: apoptosis through mitochondrial-dependent and -independent pathways. Oncogene 20:2122–2133PubMedCrossRefGoogle Scholar
  19. 19.
    Pan G, O’Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, Dixit VM (1997) The receptor for the cytotoxic ligand TRAIL. Science 276:111–113PubMedCrossRefGoogle Scholar
  20. 20.
    Walczak H, Degli-Esposti MA, Johnson RS, Smolak PJ, Waugh JY, Boiani N, Timour MS, Gerhart MJ, Schooley KA, Smith CA, Goodwin RG, Rauch CT (1997) TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL. Embo J 16:5386–5397PubMedCrossRefGoogle Scholar
  21. 21.
    Degli-Esposti MA, Smolak PJ, Walczak H, Waugh J, Huang CP, DuBose RF, Goodwin RG, Smith CA (1997) Cloning and characterization of TRAIL-R3, a novel member of the emerging TRAIL receptor family. J Exp Med 186:1165–1170PubMedCrossRefGoogle Scholar
  22. 22.
    Degli-Esposti MA, Dougall WC, Smolak PJ, Waugh JY, Smith CA, Goodwin RG (1997) The novel receptor TRAIL-R4 induces NF-kappaB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain. Immunity 7:813–820PubMedCrossRefGoogle Scholar
  23. 23.
    Srivastava RK (2001) TRAIL/Apo-2L: mechanisms and clinical applications in cancer. Neoplasia 3:535–546PubMedCrossRefGoogle Scholar
  24. 24.
    Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A (1997) Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277:818–821PubMedCrossRefGoogle Scholar
  25. 25.
    Marsters SA, Sheridan JP, Pitti RM, Huang A, Skubatch M, Baldwin D, Yuan J, Gurney A, Goddard AD, Godowski P, Ashkenazi A (1997) A novel receptor for Apo2L/TRAIL contains a truncated death domain. Curr Biol 7:1003–1006PubMedCrossRefGoogle Scholar
  26. 26.
    Shankar S, Srivastava RK (2004) Enhancement of therapeutic potential of TRAIL by cancer chemotherapy and irradiation: mechanisms and clinical implications. Drug Resist Updat 7:139–156PubMedCrossRefGoogle Scholar
  27. 27.
    Wang K, Yin XM, Chao DT, Milliman CL, Korsmeyer SJ (1996) BID: a novel BH3 domain-only death agonist. Genes Dev 10:2859–2869PubMedCrossRefGoogle Scholar
  28. 28.
    Kandasamy K, Srinivasula SM, Alnemri ES, Thompson CB, Korsmeyer SJ, Bryant JL, Srivastava RK (2003) Involvement of proapoptotic molecules Bax and Bak in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced mitochondrial disruption and apoptosis: differential regulation of cytochrome c and Smac/DIABLO release. Cancer Res 63:1712–1721PubMedGoogle Scholar
  29. 29.
    Srivastava RK, Sasaki CY, Hardwick JM, Longo DL (1999) Bcl-2-mediated drug resistance: inhibition of apoptosis by blocking nuclear factor of activated T lymphocytes (NFAT)-induced Fas ligand transcription. J Exp Med 190:253–265PubMedCrossRefGoogle Scholar
  30. 30.
    Srivastava RK, Sollott SJ, Khan L, Hansford R, Lakatta EG, Longo DL (1999) Bcl-2 and Bcl-X(L) block thapsigargin-induced nitric oxide generation, c-Jun NH(2)-terminal kinase activity, and apoptosis. Mol Cell Biol 19:5659–5674PubMedGoogle Scholar
  31. 31.
    Green DR (2005) Apoptotic pathways: ten minutes to dead. Cell 121:671–674PubMedCrossRefGoogle Scholar
  32. 32.
    Kim EJ, Suliman A, Lam A, Srivastava RK (2001) Failure of Bcl-2 to block mitochondrial dysfunction during TRAIL-induced apoptosis. Tumor necrosis-related apoptosis-inducing ligand. Int J Oncol 18:187–194PubMedGoogle Scholar
  33. 33.
    Shankar S, Chen X, Srivastava RK (2005) Effects of sequential treatments with chemotherapeutic drugs followed by TRAIL on prostate cancer in vitro and in vivo. Prostate 62:165–186PubMedCrossRefGoogle Scholar
  34. 34.
    Chen X, Thakkar H, Tyan F, Gim S, Robinson H, Lee C, Pandey SK, Nwokorie C, Onwudiwe N, Srivastava RK (2001) Constitutively active Akt is an important regulator of TRAIL sensitivity in prostate cancer. Oncogene 20:6073–6083PubMedCrossRefGoogle Scholar
  35. 35.
    Singh TR, Shankar S, Srivastava RK (2005) HDAC inhibitors enhance the apoptosis-inducing potential of TRAIL in breast carcinoma. Oncogene 24:4609–4623PubMedCrossRefGoogle Scholar
  36. 36.
    Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312PubMedCrossRefGoogle Scholar
  37. 37.
    Shankar S, Singh TR, Chen X, Thakkar H, Firnin J, Srivastava RK (2004) The sequential treatment with ionizing radiation followed by TRAIL/Apo-2L reduces tumor growth and induces apoptosis of breast tumor xenografts in nude mice. Int J Oncol 24:1133–1140PubMedGoogle Scholar
  38. 38.
    Shankar S, Singh TR, Fandy TE, Luetrakul T, Ross DD, Srivastava RK (2005) Interactive effects of histone deacetylase inhibitors and TRAIL on apoptosis in human leukemia cells: involvement of both death receptor and mitochondrial pathways. Int J Mol Med 16:1125–1138PubMedGoogle Scholar
  39. 39.
    Shankar S, Singh TR, Srivastava RK (2004) Ionizing radiation enhances the therapeutic potential of TRAIL in prostate cancer in vitro and in vivo: Intracellular mechanisms. Prostate 61:35–49PubMedCrossRefGoogle Scholar
  40. 40.
    Spierings D, McStay G, Saleh M, Bender C, Chipuk J, Maurer U, Green DR (2005) Connected to death: the (unexpurgated) mitochondrial pathway of apoptosis. Science 310:66–67PubMedCrossRefGoogle Scholar
  41. 41.
    Green DR, Kroemer G (2004) The pathophysiology of mitochondrial cell death. Science 305:626–629PubMedCrossRefGoogle Scholar
  42. 42.
    Deveraux QL, Reed JC (1999) IAP family proteins–suppressors of apoptosis. Genes Dev 13:239–252PubMedGoogle Scholar
  43. 43.
    Eiben LJ, Duckett CS (1998) The IAP family of apoptotic regulators. Results Probl Cell Differ 24:91–104PubMedGoogle Scholar
  44. 44.
    Cory S, Adams JM (2002) The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2:647–656PubMedCrossRefGoogle Scholar
  45. 45.
    Degenhardt K, Chen G, Lindsten T, White E (2002) BAX and BAK mediate p53-independent suppression of tumorigenesis. Cancer Cell 2:193–203PubMedCrossRefGoogle Scholar
  46. 46.
    Kroemer G, Reed JC (2000) Mitochondrial control of cell death. Nat Med 6:513–519PubMedCrossRefGoogle Scholar
  47. 47.
    Jacobson MD (1996) Reactive oxygen species and programmed cell death. Trends Biochem Sci 21:83–86PubMedCrossRefGoogle Scholar
  48. 48.
    Shankar S, Srivastava RK (2007) Involvement of Bcl-2 family members, phosphatidylinositol 3′-kinase/AKT and mitochondrial p53 in curcumin (diferulolylmethane)-induced apoptosis in Prostate Cancer. Inter J Oncol 30(4): 905–918Google Scholar
  49. 49.
    Azam S, Hadi N, Khan NU, Hadi SM (2004) Prooxidant property of green tea polyphenols epicatechin and epigallocatechin-3-gallate: implications for anticancer properties. Toxicol In Vitro 18:555–561PubMedCrossRefGoogle Scholar
  50. 50.
    Fujisawa S, Kadoma Y (2006) Anti- and pro-oxidant effects of oxidized quercetin, curcumin or curcumin-related compounds with thiols or ascorbate as measured by the induction period method. In Vivo 20:39–44PubMedGoogle Scholar
  51. 51.
    Korsmeyer SJ (1999) BCL-2 gene family and the regulation of programmed cell death. Cancer Res 59:1693s–1700sPubMedGoogle Scholar
  52. 52.
    Kuwana T, Newmeyer DD (2003) Bcl-2-family proteins and the role of mitochondria in apoptosis. Curr Opin Cell Biol 15:691–699PubMedCrossRefGoogle Scholar
  53. 53.
    Letai A, Bassik MC, Walensky LD, Sorcinelli MD, Weiler S, Korsmeyer SJ (2002) Distinct BH3 domains either sensitize or activate mitochondrial apoptosis, serving as prototype cancer therapeutics. Cancer Cell 2:183–192PubMedCrossRefGoogle Scholar
  54. 54.
    Murphy E, Imahashi K, Steenbergen C (2005) Bcl-2 regulation of mitochondrial energetics. Trends Cardiovasc Med 15:283–290PubMedCrossRefGoogle Scholar
  55. 55.
    Oltvai ZN, Milliman CL, Korsmeyer SJ (1993) Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609–619PubMedCrossRefGoogle Scholar
  56. 56.
    Korsmeyer SJ (1995) Regulators of cell death. Trends Genet 11:101–105PubMedCrossRefGoogle Scholar
  57. 57.
    Haldar S, Basu A, Croce CM (1997) Bcl2 is the guardian of microtubule integrity. Cancer Res 57:229–233PubMedGoogle Scholar
  58. 58.
    Levine B, Goldman JE, Jiang HH, Griffin DE, Hardwick JM (1996) Bc1-2 protects mice against fatal alphavirus encephalitis. Proc Natl Acad Sci USA 93:4810–4815PubMedCrossRefGoogle Scholar
  59. 59.
    Duchen MR (2004) Mitochondria in health and disease: perspectives on a new mitochondrial biology. Mol Aspects Med 25:365–451PubMedGoogle Scholar
  60. 60.
    Finsterer J (2004) Mitochondriopathies. Eur J Neurol 11:163–186PubMedCrossRefGoogle Scholar
  61. 61.
    Kroemer G, Zamzami N, Susin SA (1997) Mitochondrial control of apoptosis. Immunol Today 18:44–51PubMedCrossRefGoogle Scholar
  62. 62.
    Kannan K, Jain SK (2000) Oxidative stress and apoptosis. Pathophysiology 7:153–163PubMedCrossRefGoogle Scholar
  63. 63.
    Kopp P (1998) Resveratrol, a phytoestrogen found in red wine. A possible explanation for the conundrum of the ‘French paradox’? Eur J Endocrinol 138:619–620PubMedCrossRefGoogle Scholar
  64. 64.
    Bhat KP, Lantvit D, Christov K, Mehta RG, Moon RC, Pezzuto JM (2001) Estrogenic and antiestrogenic properties of resveratrol in mammary tumor models. Cancer Res 61:7456–7463PubMedGoogle Scholar
  65. 65.
    Jang DS, Kang BS, Ryu SY, Chang IM, Min KR, Kim Y (1999) Inhibitory effects of resveratrol analogs on unopsonized zymosan-induced oxygen radical production. Biochem Pharmacol 57:705–712PubMedCrossRefGoogle Scholar
  66. 66.
    Gescher AJ, Steward WP (2003) Relationship between mechanisms, bioavailibility, and preclinical chemopreventive efficacy of resveratrol: a conundrum. Cancer Epidemiol Biomarkers Prev 12:953–957PubMedGoogle Scholar
  67. 67.
    Walle T, Hsieh F, DeLegge MH, Oatis JE Jr, Walle UK (2004) High absorption but very low bioavailability of oral resveratrol in humans. Drug Metab Dispos 32:1377–1382PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Sharmila Shankar
    • 1
  • Imtiaz Siddiqui
    • 1
  • Rakesh K. Srivastava
    • 1
  1. 1.Department of BiochemistryThe University of Texas Health Science Center at TylerTylerUSA

Personalised recommendations