Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Induction of Cdk inhibitor p21 and inhibition of cyclooxygenase-2 by resveratrol in human lung carcinoma A549 cells.

Resveratrol에 의한 A549 인체 폐암세포의 증식억제 및 apoptosis 유발에 관한 연구

  • 김영애 (일본 부산의과약과대학 화한약연구소) ;
  • 임선영 (한국해양대학교 해양과학기술대학 해양과학부) ;
  • 이숙희 (부산대학교 생활환경대학 식품영양학과) ;
  • 박건영 (부산대학교 생활환경대학 식품영양학과) ;
  • 이원호 (부산대학교 자연과학대학 생물학과) ;
  • 최영현 (동의대학교 한의과대학 생화학교실)
  • Published : 2004.10.01
Download PDF
⟨ Previous Next ⟩

Abstract

Resveratrol, a phytoalexin found at high levels in grapes and in grape products such as red wine, has been reported to possess a wide range of biological and pharmacological activities including antioxident, anti-inflammatory, anti-mutagenic, and anti-carcinogenic effects. According to recent studies, this compound is an effective inhibitor of cell growth in general, triggers partial arrest of the cell cycle and induce apoptosis. In this study, the anti-proliferative effects of resveratrol in A549 human lung carcinoma cells were investigated. It is shown that resveratrol induced the growth inhibition in a time-dependent manner and morphological changes of A549 cells, which were associated with induction of S phase arrest of the cell cycle and apoptotic cell death. The Bcl-$X_L$levels were markedly down-regulated in resveratrol treated cells, however, Bax and Bcl-2 were remained unchanged. Resveratrol treatment induced the proteolytic degradation of Sp-l and proliferating cell nuclear antigen protein, and inhibited the expression of $\beta$-catenin protein. Resveratrol treatment also induced a marked up-regulation of cyclin-dependent kinase (Cdk) inhibitor p21 and inhibited the kinase activities of Cdk2 and Cdk4. In addition, resveratrol treatment inhibited the levels of cyclooxygenase (COX)-2 mRNA and protein, and the release of prostagladin E2 without alteration of COX-1 expression. Taken together, these findings suggest that resveratrol may be a potential chemotherapeutic agent for the control of human lung carcinorma cells.

Resveratrol은 포도와 같은 식물에서 각종 감염균으로부터 자신의 몸을 보호하기 위하여 생성되는 물질인 phytoalexin의 일종으로 강력한 항산화작용, 암예방 효과 및 항암 작용을 포함한 각종 약리작용을 가진 것으로 보고되어져 오고 있다. 본 연구에서는 resveratrol의 항암작용 기전해석을 위하여 A549 인체폐암세포의 종식에 미치는 resverakol의 영향을 조사하였다. A549 세포의 생존율은 resveratrol의 처리시간 증가에 따라 강력하게 억제되었으며, 이는 암세포의 다양한 형태변형을 동반한 세포주기 C2/M arrest 및 염색질 응축 현상을 동반한 apoptosis 유발에 의한 것임을 알 수 있었다. Resveratrol 처리에 의한 apoptosis 유발은 Bcl-2의 발현변화 없이 Bcl-$X_L$의 발현 감소에 따른 상대적인 Bax의 발현 증가와 Sp-1, PCNA 및 $\beta$-catenin 등과 같은 단백질의 분해 현상과 연관성이 있었다 또한 resveratrol에 의한 A549세포 의 증식억제는 Cdk inhibitor p21의 발현 증가에 따른 Cdks 의 kinase 활성 저하 및 COX-2의 선택적 저해에 따른 PGE2 생성 저하와 관련이 있었다.

Keywords

References

  1. Ahmad, N., V.M. Adhami, F. Afaq, D.K. Feyes and H. Mukhtar. 2001. Reveratrol 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
  2. Arends, M.J., R.G. Morris and A.H. Wyllie. 1990. Apoptosis. The role of the endonuclease. Am J Pathol 136, 593-608
  3. Arichi, H., Y. Kimura, H. Okuda, K. Baba, M. Kozawa and S. Arichi. 1982. Effects of stilbene components of the roots of Polygonum cuspidatum Sieb. et Zucc. on lipid metabolism. Chem Pharm Bull 30, 1766-1770 https://doi.org/10.1248/cpb.30.1766
  4. Barry, M.A., C.A. Behnke and A. Eastman. 1990. Activation of programmed cell death (apoptosis) by cisplatin, other anticancer drugs, toxins and hyperthermia. Biochem Pharmacol 40, 2353-2362 https://doi.org/10.1016/0006-2952(90)90733-2
  5. Brancolini, C., D. Lazarevic, J. Rodriguez and C. Schneider. 1997. Dismantling cell-cell contacts during apoptosis is coupled to a caspase-dependent proteolytic cleavage of $\beta$- catenin. J Cell Biol 139, 759-771 https://doi.org/10.1083/jcb.139.3.759
  6. Chiarugi, V., L. Magnelli and G. Basi. 1994. Apoptosis and the cell cycle. Cell Mol Biol Res 40, 603-612
  7. Chiarugi, V., L. Magnelli and O. Gallo. 1998. Cox-2, iNOS and p53 as play-makers of tumor angiogenesis. Int J Mol Med 2, 715-719
  8. Choi, Y.H., W.H. Lee, L. Zhang and K.Y. Park. 2000. p53- independent induction of $p21^{WAF1/CIP1}$, reduction of cyclin B1 and G2/M arrest by the isoflavones genistein in human prostate carcinoma cells. Jpn J Cancer Res 91, 164-173 https://doi.org/10.1111/j.1349-7006.2000.tb00928.x
  9. Choi, Y.H., K.R. Kong, Y.A. Kim, K.O. Jung, J.H. Kil, S.H. Rhee and K.Y. Park. 2003. Induction of Bax and activation of caspases during $\beta$-sitosterol-mediated apoptosis in human colon cancer cells, Int J Oncol 23, 1657-1661
  10. Clement, M.V., J.L. Hirpara, CS.H. Hawdhury and S. Pervaiz. 1998. Chemopreventive agent resveratrol, a natural product derived from grapes, triggers CD95 signaling- dependent apoptosis in human tumor cells. Blood 92, 996- 1002
  11. Dempke, W., C. Rie, A. Grothey and H.J. Schmoll. 2001. Cyclooxygenase-2: a novel target for cancer chemotherapy? J Cancer Res Clin Oncol 127, 411-417 https://doi.org/10.1007/s004320000225
  12. Dercks, W. and L.L. Creasy. 1989. The significance of stilbene phytoalexins in the Plasmopara viticola-grapevine interaction. Physiol Mol Plant Pathol 34, 189-202 https://doi.org/10.1016/0885-5765(89)90043-X
  13. El-Deiry, W.S., T. Tokino, V.E. Velculesco, D.B. Levy, R. Parsons, J.M. Trent, D. Lin, D., E.W. Mercer, K.W. Kinzler and B. Vogelstain. 1993. WAF1, a potential mediator of p53 tumor suppression. Cell 75, 817-825 https://doi.org/10.1016/0092-8674(93)90500-P
  14. Estrov, Z., S. Shishodia, S. Faderl, D. Harris, Q. Van, H.M. Kantarjian, M. Talpaz and B.B. Aggarwal. 2003. Resveratrol blocks interleukin-1$\beta$-induced activation of the nuclear transcription factor NF-$_{k}$B, inhibits proliferation, causes S-phase arrest, and induces apoptosis of acute myeloid leukemia cells. Blood 102, 987-995 https://doi.org/10.1182/blood-2002-11-3550
  15. Evans, V.G. 1995. Multiple pathways to apoptosis. Cell Biol Int 17, 461-476 https://doi.org/10.1006/cbir.1993.1087
  16. Fesus, L., Z. Szondy and I. Uray. 1995. Probing the molecular program of apoptosis by cancer chemopreventative agents. J Cell Biochem 22, 151-161 https://doi.org/10.1002/jcb.240590820
  17. Giercksky, K.E. 2001. COX-2 inhibition and prevention of cancer. Best Pract Res Clin Gastroenterol 15, 821-833 https://doi.org/10.1053/bega.2001.0237
  18. Goldberg, D.M., S.E. Hahn and J.G. Parkes. 1995. Beyond alcohol-beverage consumption and cardiovascular mortality. Clin Chim Acta 237, 155-187 https://doi.org/10.1016/0009-8981(95)06069-P
  19. Hickman, J.A. 1992. Apoptosis induced by anticancer drugs. Cancer Metastasis Rev 11, 121-129 https://doi.org/10.1007/BF00048059
  20. Hockenbery, D.M., Ζ.Ν. Oltvai, Χ.Μ. Yin, C.L. Milliman and S.J. Korsmeyer. 1993. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75, 241-251 https://doi.org/10.1016/0092-8674(93)80066-N
  21. Huang, C., W.Y. Ma, A. Goranson and Z. Dong. 1999. Resveratrol suppresses cell transformation and induces apoptosis through a p53-dependent pathway. Carcinogenesis 20, 237-242 https://doi.org/10.1093/carcin/20.2.237
  22. Huang, Y., S. Nakada, T. Ishiko, T. Utsugisawa, R. Datta, S. Kharbanda, K. Yoshida, R.V. Talanian, R. Weichselbaum, D. Kufe and Z.M. Yuan. 1999. Role for caspase-mediated cleavage of Rad51 in induction of apoptosis by DNA damage. Mol Cell Biol 19, 2986-2997
  23. Jang, J.H. and Y.J. Surh. 2003. Protective effect of resveratrol on $\beta$-amyloid-induced oxidative PC12 cell death. Free Radic BioI Med 34, 1100-1110 https://doi.org/10.1016/S0891-5849(03)00062-5
  24. Jang, J.H. and Y.J. Surh. 2001. Protective effects of resveratrol on hydrogen peroxide-induced apoptosis in rat pheochromocytoma (PCI2) cells. Mutat Res 496, 181-190 https://doi.org/10.1016/S0891-5849(03)00062-5
  25. Jang, M., L. Cai, G.O. Udeani, K.V. Slowing, C.F. Thomas, C.W.W. Beecher, H.H.S. Fong, N.R. Farnsworth, A.D. Kinghorn, R.G. Mehta, R.C. Moon and J.M. Pezzuto. 1997. Cancer chemopreventative activity of resveratrol, a natural product derived from grapes. Science 275, 218-220 https://doi.org/10.1126/science.275.5297.218
  26. Jo, E.H., H.D. Hong, N.C. Ahn, J.W. Jung, S.R. Yang, J.S. Park, S.H. Kim, Y.S. Lee and K.S. Kang. 2004. Modulations of the Bcl-2/Bax family were involved in the chemopreventive effects of licorice root (Glycyrrhiza uralensis Fisch) in MCF-7 human breast cancer cell. J Agrie Food Chem 52, 1715-1719 https://doi.org/10.1021/jf035012t
  27. Joe, A.K., H. Liu, M. Suzui, M.E. Vural, D. Xiao and I.B. Weinstein. 2002. Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines. Clin Cancer Res 8, 893-903
  28. Kim, YA, B.T. Choi, Y.T. Lee, D.I. Park, S.H. Rhee, K.Y. Park and Y.H. Choi. 2004. Resveratrol inhibits cell proliferation and induces apoptosis of human breast carcinoma MCF-7 cells. Oneal Rep 11, 441-446
  29. Kim, Y.A., W.H. Lee, T.H. Choi, S.H. Lee, K.Y. Park and Y.H. Choi. 2003. Involvement of p21, pRB, Bax and NF-$_{k}$B in induction of growth arrest and apoptosis by resveratrol in human lung carcinoma A549 cells. Int J Oneal 23, 1143-1149
  30. Larrosa, M., F.A. Tomas-Barberan and J.C. Espin. 2003. Grape polyphenol resveratrol and the related molecule 4-hydroxystilbene induce growth jnhibition, apoptosis, S-phase arrest, and upregulation of cyclins A, E, and B1 in human SK-Mel-28 melanoma cells. J Agric Food Chem 51, 4576-4584 https://doi.org/10.1021/jf030073c
  31. Liu, W.K., S.X. Xu and C.T. Che. 2000. Anti-proliferative effect of ginseng saponins on human prostate cancer cell line. Life Sci 67, 1297-1306 https://doi.org/10.1016/S0024-3205(00)00720-7
  32. Lowe, S.W., H.E. Ruley, T. Jacks and D.E. Housman. 1993. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74, 957-967 https://doi.org/10.1016/0092-8674(93)90719-7
  33. Miyashita, T. and J.C Reed. 1995. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80, 293-299 https://doi.org/10.1016/0092-8674(95)90412-3
  34. Morgan, D.O. 1995. Principles of CDK regulation. Nature 374, 131-134 https://doi.org/10.1038/374131a0
  35. Paceasciak, C.R., S. Hahn, E.P. Diamandis, G. Soleas and D.M. Goldberg. 1995. The red wine phenolics transresveratrol and quercetin block human platelet aggregation and eicosanoid synthesis-implications for protection against coronary heart disease. Clin Chim Acta 235, 207-219 https://doi.org/10.1016/0009-8981(95)06045-1
  36. Reddy, B.S., C.X. Wang, H. Samaha, R. Lubet, V.E. Steele, G.J. Kelloff and C.V. Rao. 1997. Chemoprevention of colon carcinogenesis by dietary perillyl alcohol. Cancer Res 57, 420-425
  37. Reed, J.C. 1998. Bcl-2 family proteins, Oncogene 17, 3225- 3236 https://doi.org/10.1038/sj.onc.1202591
  38. Rickers, A., N. Peters, V. Badock, R. Beyaert, P. Vandenabeele, B. Dorken and, K. Bommert. 1999. Cleavage of transcription factor SP1 by caspases during anti-IgM- induced B-cell apoptosis. Eur J Biochem 261, 269-274 https://doi.org/10.1046/j.1432-1327.1999.00273.x
  39. She, Q.B., A.M. Bode, W.Y. Ma, N.Y. Chen and Z. Dong. 2001. Resveratrol-induced activation of p53 and apoptosis is mediated by extracellular-signal-regulated protein kinases and p38 kinase. Cancer Res 61, 1604-1610
  40. Sherr, C.J. 2000. The Pezcoller lecture: cancer cell cycles revisited. Cancer Res 60, 3689-3695
  41. Subbaramaiah, K., W.J. Chung, P. Michaluart, N. Telang, T. Tanabe, H. Inoue, M. Jang, J.M. Pezzuto and A.J. Danneberg, Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary dpithelial cells. J Biol Chem 273, 21875-21882 https://doi.org/10.1074/jbc.273.34.21875
  42. Surh, Y.J., Y.J. Hurh, J.Y. Kang, E. Lee, G. Kong and S.J. Lee. 1999. Resveratrol, an antioxidant present in red wine, induces apoptosis in human promyelocytic leukemia (HL- 60) cells. Cancer Lett 140, 1-10 https://doi.org/10.1016/S0304-3835(99)00039-7
  43. Surh, Y.J., K.S. Chun, H.H. Cha, S.S. Han, Y.S. Keum, K.K. Park and S.S. Lee. 2001. S.S., Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-$_{k}$B activation. Mutat Res 480- 481, 243-268 https://doi.org/10.1016/S0027-5107(01)00183-X
  44. Tewari, M., L.T. Quan, K. O'Rourke, S. Desnoyers, Z. Zeng, D.R. Beidler, G.G. Poirier, G.S. Salvesen and V.M. Dixit. 1995. Yama/CPP32 $\beta$, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell 81, 801- 809 https://doi.org/10.1016/0092-8674(95)90541-3
  45. Tsan, M.F., J.E. White, J.G. Maheshwari and G. Chikkappa. 2002. Anti-leukemia effect of resveratrol. Leuk Lymphoma 43, 983-987
  46. Turini, M.E. and R.N. DuBois. 2002. Cyclooxygenase-2: a therapeutic target. Annu Rev Med 53, 35-57 https://doi.org/10.1146/annurev.med.53.082901.103952
  47. Vainio, H. 2001.Is COX-2 inhibition a panacea for cancer prevention? Int J Cancer 94, 613-614 https://doi.org/10.1002/ijc.1518
  48. Wolter, F., B. Akoglu, A. Clausnitzer and J. Stein. 2001. Downregulation of the cyclin D1/Cdk4 complex occurs during resveratrol-induced cell cycle arrest in colon cancer cell lines. J Nutr 131, 2197-2203

Cited by

  1. A Study on Peanut Spouts Extract as the Anti-oxidant Activity and the Skin Whitening Cosmetic Ingredients vol.31, pp.1, 2016, https://doi.org/10.7841/ksbbj.2016.31.1.14