DOI QR코드

DOI QR Code

Resveratrol Induces Apoptosis through PI3K/Akt and p53 Signal Pathway in MDA-MB-231 Breast Cancer Cells

Resveratrol이 MDA-MB-231 유방암 세포에서 PI3K/Akt와 p53 신호경로를 통한 apoptosis 유도

  • Kwon, Jung-Ki (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Park, Young-Seok (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Park, Byung-Kwon (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, Byeong-Soo (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, Sang-Ki (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Jung, Ji-Youn (Department of Companion and Laboratory Animal Science, Kongju National University)
  • Received : 2012.05.31
  • Accepted : 2012.06.15
  • Published : 2012.08.31

Abstract

This study was conducted in order to investigate the effect of resveratrol on the induction of apoptosis in MDA-MB-231 breast cancer cells. The result of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT) assay shows that cell viability significantly decreased in a dose and time-dependent manner. 4',6-diamidino-2-phenylindole (DAPI) staining shows significantly increased chromatin condensation in a dose and time-dependent manner. Resveratrol increased the expression of p53, cleaved-caspase-3, and cleaved-caspase-9, whereas the expression of PI3K/Akt decreased in a time-dependent manner. We investigated the in vivo tumor growth inhibitory effect of resveratrol. Tumor volume was significantly decreased in the 50 mg/kg resveratrol-administration group compared to the control group. In the 50 mg/kg treated group. Apoptosis cells were frequently observed by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay. Immunohistochemistry staining shows increased the expression of p53, cytochrome-C, and cleaved-caspase-3 in the 50 mg/kg treated group. These results indicate that resveratrol induced apoptosis through PI3K/Akt and p53 signal pathway in MDA-MB-231 cell.

본 연구는 MDA-MB-231 세포에서 resveratrol의 apoptosis 유발 효과에 대해 알아보기 위해 연구되었다. Cell viability 결과 농도 유의적으로 감소하였으며, DAPI stain에서는 농도 의존적으로 chromatin condensation이 증가하는 것을 확인하였다. Resveratrol은 p53, cleaved-caspase-3, cleaved-caspase-9의 발현을 증가시켰지만, PI3K/Akt의 발현은 시간 의존적으로 감소시켰다. In vivo 실험에서는 resveratrol의 종양 억제 효과를 확인하였다. 50 mg/kg 투여한 군에서 종양의 크기가 대조군에 비해 감소하였으며, TUNEL assay를 통해 apoptosis cell을 관찰한 결과 50 mg/kg 투여한 군에서 많이 관찰되었다. 면역조직화학 염색을 통해 50 mg/kg 투여한 군에서 p53, cytochrome-C, cleaved-caspase-3의 발현이 증가하는 것을 확인하였다. 본 연구의 결과를 종합하여 봤을 때 resveratrol이 MDA-MB-231 세포에 apoptosis를 유발하는데 효과가 있는 것으로 사료된다.

Keywords

References

  1. Chung GS, Kim HD. Improvement and analysis of fire-fighting service in an aging society. J. Korean Inst. Fire Sci. Eng. 25: 27-33 (2011)
  2. World cancer research fund international. Available from: http://www.wcrf.org/cancer_statistics/world_cancer_statistics.php. Accessed May 30, 2012.
  3. National cancer center. Available from: http://www.cancer.go.kr/ncic/cics_b/01/013/1268116_5873.html. Accessed May 30, 2012.
  4. Yoo GS, Lee JM, Lee CH, Jang JB, Lee KS. Study of apoptosis by scirpi tuber in Hela cell and MCF-7 cell. J. Oriental Obstet. Gynecol. 24: 1-13 (2011)
  5. Choi DY. Effect of growth inhibition in Hep3B cell and HeLa cell by treatment of Euonymus alatus(Thunb.) Sieb extracts. Dongguk J. Inst. Oriental Med. 7: 155-162 (1999)
  6. Jeong KA, Park KM, Cho SH. Anti-proliferative effects of samneung (Sparganii Rhizoma) extract on MCF-7 cells. J. Oriental Obstet. Gynecol. 19: 166-177 (2006)
  7. Park YS, Han JY, Lee TK, Kim DI. Growth inhibitory and antioxidative effects of crude methanolic extract from Euonymus alatus (Thunb.) Sieb on SKBR3 human breast cancer cell line. J. Oriental Obstet. Gynecol. 18: 45-54 (2005)
  8. Yong HS, Ko SG. Inhibition of cellular proliferation and apoptosis by Scutellaria Baicalensis in MDA-MB-231 breast cancer cells. Korean J. Orient. Int. Med. 25: 451-460 (2004)
  9. Lee JS, Lee SH, Jang YM, Lee JD, Lee BH, Jung JY. Macrophage and anticancer activities of feed additives on $\beta$-glucan from Schizophyllum commune in breast cancer cells. J. Korean Soc. Food. Sci. Nutr. 40: 949-955 (2011) https://doi.org/10.3746/jkfn.2011.40.7.949
  10. Woo EY, Park SY, Kwon SJ, Kwon GT, Kim JD, Lim SS, Yoon JH. Effect of eupatorium japonicum extract on the metastasis, invasion, and adhesion of MDA-MB-231 human breast cancer cells. Korean J. Food. Sci. Technol. 43: 213-219 (2011) https://doi.org/10.9721/KJFST.2011.43.2.213
  11. Feig SA. Effect of service screening mammography on population mortality from breast carcinoma. Cancer 95: 451-457 (2002) https://doi.org/10.1002/cncr.10764
  12. Tabar L, Yen MF, Vitak B, Chen HH, Smith RA, Duffy SW. Mammography service screening and mortality in breast cancer patients: 20-year follow-up before and after introduction of screening. Lancet 361: 1405-1410 (2003) https://doi.org/10.1016/S0140-6736(03)13143-1
  13. Zhou J, Zhong Y. Breast cancer immunotherapy. Cell. Mol. Immunol. 1: 247-255 (2004)
  14. Liu RH. Potential synergy of phytochemicals in cancer prevention: Mechanism of action. J. Nutr. 134: 3479S-3485S (2004) https://doi.org/10.1093/jn/134.12.3479S
  15. Cho CH, Kim SY, Yoo GJ, Son MH, Park KH, Lim BL, Kim DC, Chae HJ. Resveratrol extraction from grape fruit stem and its antioxidant activity. J. Korean Soc. Appl. Biol. Chem. 51: 11-16 (2008)
  16. Gorham J. The stilbenoids. Prog. Phytochem. 6: 203-209 (1980)
  17. Kim DH, Heo TH, Kim JB, Kim SJ. Resveratrol upregulates p21 cell cycle regulator, in gaucher disease cells. J. Life Sci. 20: 1281-1286 (2010) https://doi.org/10.5352/JLS.2010.20.8.1281
  18. Constant J. Alcohol, ischemic heart disease, and the french paradox. Clin. Cardiol. 20: 420-424 (1997) https://doi.org/10.1002/clc.4960200504
  19. Fauconneau B, Waffo-Tegu P, Huguet F, Barrier L, Decendit A, Merillon JM. Comparative study of radical scavenger and antioxidant properties of phenolic compounds from vitis vinifera cell cultures using in vitro tests. Life Sci. 61: 2103-2110 (1997) https://doi.org/10.1016/S0024-3205(97)00883-7
  20. Pace-Asciak CR, Hahn SE, Diamandis EP, Soleas G, Goldberg DM. The red wine phenolics trans-resveratrol and quercetin block human platelet aggregation and eicosanoid synthesis: Implications for protection against coronary heart disease. Clin. Chim. Acta 235: 207-219 (1995) https://doi.org/10.1016/0009-8981(95)06045-1
  21. Lee HS, Sur EY, Kim WK. Resveratrol induces apoptosis in SW480 human colon cancer cell lines. Food Sci. Biotechnol. 13: 80-84 (2004)
  22. Jang M, Ca L, Udeani GO, Slowing KV, Thomas CF, Beecher CWW, Fong HHS, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275: 218-220 (1997) https://doi.org/10.1126/science.275.5297.218
  23. Guengerich FP. Roles of cytochrome P-450 enzymes in chemical carcinogenesis and cancer chemotherapy. Cancer Res. 48: 2946-2954 (1988)
  24. Jiang H, Zhang L, Kuo J, Kuo K, Gautam SC, Groc L, Rodriguez AI, Koubi D, Hunter TJ, Corcoran GB, Seidman MD, Levine RA. Resveratrol-induced apoptotic death in human U251 glioma cells. Mol. Cancer Ther. 4: 554-561 (2005) https://doi.org/10.1158/1535-7163.MCT-04-0056
  25. Aziz MH, Nihal M, Fu VX, Jarrard DF, Ahmad N. Resveratrolcaused apoptosis of human prostate carcinoma LNCaP cells is mediated via modulation of phosphatidylinositol 3'-kinase/Akt pathway and Bcl-2 family proteins. Mol. Cancer Ther. 5: 1335-1341 (2006) https://doi.org/10.1158/1535-7163.MCT-05-0526
  26. Vanamala J, Reddivari L, Radhakrishnan S, Tarver C. Resveratrol suppresses IGF-1 induced human colon cancer cell proliferation and elevates apoptosis via suppression of IGF-1R/Wnt and activation of p53 signaling pathways. BMC Cancer 10: 238 (2010) https://doi.org/10.1186/1471-2407-10-238
  27. Carnero A. The PKB/AKT pathway in cancer. Curr. Pharm. Des. 16: 34-44 (2010) https://doi.org/10.2174/138161210789941865
  28. Osaki M, Oshimura M, Ito H. PI3K-Akt pathway: Its functions and alterations in human cancer. Apoptosis 9: 667-676 (2004) https://doi.org/10.1023/B:APPT.0000045801.15585.dd
  29. Amaral JD, Xavier JM, Steer CJ, Rodrigues CM. The role of p53 in apoptosis. Discov. Med. 9: 145-152 (2010)
  30. Yonish-Rouach E, Resnitzky D, Lotem J, Sachs L, Kimchi A, Oren M. Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6. Nature 352: 345-347 (1991) https://doi.org/10.1038/352345a0
  31. Lowe SW, Schmitt EM, Smith SW, Osborne BA, Jacks T. p53 is required for radition-induced apoptosis in mouse thymocyte. Nature 362: 847-849 (1993) https://doi.org/10.1038/362847a0
  32. Choi WJ, Youn SH, Back JH, Park S, Park EJ, Kim KJ, Park HR, Kim AL, Kim KH. The role of KLF4 in UVB-induced murine skin tumor development and its correlation with cyclin D1, p53, and p21(Waf1/Cip1) in epithelial tumors of the human skin. Arch. Dermatol. Res. 303: 191-200 (2011) https://doi.org/10.1007/s00403-010-1101-0
  33. Millau JF, Bandele OJ, Perron J, Bastien N, Bouchard EF, Gaudreau L, Bell DA, Drouin R. Formation of stress-specific p53 binding patterns is influenced by chromatin but not by modulation of p53 binding affinity to response elements. Nucleic Acids Res. 39: 3053-3063 (2010)
  34. Sheu MJ, Chou PY, Huang CS, Tsai IC, Chien YC, Lin SY, Tsai HY, Cheng HC, Wu CH. Pipoxolan inhibits proliferation of HL-60 human leukaemia cancer cells by arresting the cell cycle at the G0/G1 phase. Clin. Exp. Pharmacol. Physiol. 37: 605-612 (2010) https://doi.org/10.1111/j.1440-1681.2010.05358.x
  35. Casanova F, Quarti J, da Costa DC, Ramos CA, da Silva JL, Fialho E. Resveratrol chemosensitizes breast cancer cells to melphalan by cell cycle arrest. J. Cell. Biochem. 113: 2586-2596 (2012) https://doi.org/10.1002/jcb.24134
  36. Dulic V, Kaufmann WK, Wilson SJ, Tisty TD, Lees E, Harper JW, Elledge SJ, Reed SI. p53 -dependent inhibition of cyclindependent kinase activities in human fibroblasts during radiationinduced $G_{1}$ arrest. Cell 76: 1013-1023 (1994) https://doi.org/10.1016/0092-8674(94)90379-4
  37. El-Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, Pietenpol JA, Burrell M, Hill DE, Wang Y, Wiman KG, Mercer WE, Kastan MB, Kohn KW, Elledge SJ, Kinzler KW, Vogelstein B. WAF1/CIP1 is induced in p53-mediated $G_{1}$ arrest and apoptosis. Cancer Res. 54: 1169-1174 (1994)
  38. Oda E, Ohki R, Murasawa H, Nemoto J, Shibue T, Yamashita T, Tokino T, Taniguchi T, Tamaka N. Noxa, a BH-3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis. Science 288: 1053-1058 (2000) https://doi.org/10.1126/science.288.5468.1053
  39. Nakano K, Vousden KH. PUMA, a novel proapoptotic gene, is induced by p53. Mol. Cell 7: 683-694 (2001) https://doi.org/10.1016/S1097-2765(01)00214-3
  40. Park SY, Kim IS, Lee SH, Lee SH, Jung DW, Park OJ, Kim YM. Anti-proliferative effects of selenium in HT-29 colon cancer cells via inhibition of Akt. J. Life Sci. 22: 55-61 (2012) https://doi.org/10.5352/JLS.2012.22.1.55

Cited by

  1. Induction of Apoptosis by Piceatannol in YD-15 Human Oral Cancer Cells vol.44, pp.7, 2015, https://doi.org/10.3746/jkfn.2015.44.7.975
  2. Extracellular NAMPT/visfatin causes p53 deacetylation via NAD production and SIRT1 activation in breast cancer cells vol.35, pp.6, 2017, https://doi.org/10.1002/cbf.3279