Annals of Surgical Treatment and Research

The Korean Surgical Society (대한외과학회)
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Role of p53 and p38 MAPK on Doxorubicin and Lovastatin-induced Apoptosis in Colon Cancer Cells

대장암 세포주에서 Doxorubicin과 Lovastatin 유도성 세포고사에 대한 p53과 p38 MAPK의 영향

Park, Won-Cheol;Lee, Jeong-Kyun;Park, Rae-Kil
박원철;이정균;박래길

  • Published : 20030000
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Abstract

Purpose: Apoptosis plays a central role in tumor development and it has been hypothesized that lack or failure of apoptosis leads to the development of tumors, including colon cancers. Anticancer drugs do not invariably cytotoxic to all cancer cells. Some resistant cells against anticancer drugs are resuming to proliferate after initial damage, whereas others undergo permanent growth arrest or death. The resistance of advanced colorectal cancers to chemotherapy is often related to mutations in p53 tumor suppressor gene. Methods: To understand the molecular mechanism of lovastatin-induced apoptosis, the relationship between p53 and p38 MAPK upon cytotoxicity by anticancer drug was investigated in colon cancer cell lines including HCT-116 (p53 wild type) and HT-29 (mutated p53). Doxorubicin (dox) or lovastatin (lova) increased the cytotoxicity of colon cancer cells in a dose- and time-dependent manner. Results: Cytotoxic effects of dox and lova was more prominent in HCT-116 than HT-29. The combined treatment of dox and lova further increased the cytotoxic effect on both cells. The cytotoxicity of dox and lova was resulted from apoptotic cell death, which was determined by genomic DNA fragmentation, PARP cleavage and activation of caspase-3 and -9. The Combined treatment of dox and lova synergistically increased the catalytic activity of caspase-3 and -9 in colon cancer cells, but not caspase-6 and -8. Anticancer drugs also induced the cytosolic release of cytochrome c from mitochondria. Dox also induced the accumulation and activation of p53 protein (phosphorylation of p53, ser15), which was suppressed by lova. p38 MAPK inhibitor, SB203580, significantly increased the apoptotic death of lova-treated cells. Furthermore, Lova significantly induced apoptotic death of MKK6 D/D stable transfectant compared to pcDNA3.1 transfectant of HT-29 cells, which was determined by MTT assay, DNA fragmentation, and caspase activation. Similarly to MKK6 D/D transfectant, Exip (alternative splicing p38 gene) transfectant showed the apoptogenic effect of dox with lova. Conclusion: These results indicate that lova induces apoptosis in colon cancer cells via p38 MAPK-dependent and p53-independent mechanism.

Keywords

References

  1. Roth JA, Nguyen D, Lawrence DD, Kemp BL, Carrasco CH, Ferson DZ, et al. Retrovirus-mediated wild-type p53 gene transfer to tumors of patients with lung cancer. Nat Med 1996;2:985-91 https://doi.org/10.1038/nm0996-985
  2. Conseiller E, Debussche L, Landais D, Venot C, Maratrat M, Sierra V, et al. CTS1: a p53-derived chimeric tumor suppressor gene with enhanced in vitro apoptotic properties. J Clin Invest 1998;101:120-7 https://doi.org/10.1172/JCI1140
  3. Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 1995;270:1326-311 https://doi.org/10.1126/science.270.5240.1326
  4. Denizat F, Lang R. Rapid colorimetric assay for cell growth and survival. Modification the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 1986;89:271-7 https://doi.org/10.1016/0022-1759(86)90368-6
  5. Ferreira CG, Span SW, Peters GJ, Kruyt FA, Giaccone G. Chemotherapy triggers apoptosis in a caspase-8-dependent and mitochondria-controlled manner in the non small cell lung cancer cell line NCI-H460. Cancer Res 2000;60:7133-41
  6. Sudo T, Yagasaki Y, Hama H, Watanabe N, Osada H. Exip, a new alternative splicing variant of p38 alpha, can induce an earlier onset of apoptosis in HeLa cells. Biochem Biophys Res Commun 2002;291:838-43 https://doi.org/10.1006/bbrc.2002.6529
  7. Padayatty SJ, Marcelli M, Shao TC, Cunningham GR. Lovastatin induced apoptosis in prostate stromal cells. J Clin Endocrinol Metab 1997;82:1434-9 https://doi.org/10.1210/jc.82.5.1434
  8. Reedquist KA, Pope TK, Roess DA. Lovastatin inhibits proliferation and differentiation and causes apoptosis in lipopolysaccharide-stimulated murine B cells. Biochem Biophys Res Commun 1995;211:665-70 https://doi.org/10.1006/bbrc.1995.1863
  9. Perez-Sala D, Mollinedo F. Inhibition of ioprenoid biosynthesis induces apoptosis in human promyelocytic HL-60 cells. Biochem Biophys Res Commun 1994;199:1209-15 https://doi.org/10.1006/bbrc.1994.1359
  10. Satoh S, Isobe H, Ayukawa K, Sakai H, Nawata H. The effects of pravastatin, and HMG-CoA reductase inhibitor, on cell viability and DNA production of rat hepatocytes. Life Sci 1996;59:1103-8 https://doi.org/10.1016/0024-3205(96)00427-4
  11. Agarwal B, Bhendwal S, Halmos B, Moss SF, Ramey WR, Holt PR. Lovastatin augments apoptosis induced by chemotherapeutic agents in colon cancer cells. Clin Cancer Res 1999;5:2223-9
  12. Feleszko W, Jakobisiak M. Lovastatin augments apoptosis induced by chemotherapeutic agents in colon cancer cells. Clin Cancer Res 2000;6:1198-9
  13. Kastan MB, Zhan Q, el Deiry WS, Carrier F, Jacks T, Walsh WV, et al. A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 1992;71:587-97 https://doi.org/10.1016/0092-8674(92)90593-2
  14. Fritsche M, Haessler C, Brandner G. Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents. Oncogene 1993;8:307-18
  15. Maltzman W, Czyzyk L. UV irradiation stimulates levels of p53 cellular tumor antigen in nontransformed mouse cells. Mol Cell Biol 1994;4:1689-94
  16. el-Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, et al. WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 1994;54:1169-74
  17. El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, et al. WAF1, a potential mediator of p53 tumor suppression. Cell 1993;19:817-25 https://doi.org/10.1016/0092-8674(93)90500-P
  18. Tanaka H, Arakawa H, Yamaguchi T, Shiraish K, Fukuda S, Matsui K, et al. A ribonucleotide reductase gene involved in a p53-dependent cell-cycle checkpoint for DNA damage. Nature 2000;404:42-9 https://doi.org/10.1038/35003506
  19. Miyashita, T, Reed JC. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 1995;80:293-9 https://doi.org/10.1016/0092-8674(95)90412-3
  20. Yin C, Knudson CM, Korsmeyer SJ, Van Dyke T. Bax suppresses tumorigenesis and stimulates apoptosis in vivo. Nature 1997;385:637-640 https://doi.org/10.1038/385637a0
  21. Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B. A model for p53-induced apoptosis. Nature 1997;389:300-5 https://doi.org/10.1038/38525
  22. Bennett M, MacDonald K, Chan SW, Luzio JP, Simari R, Weissberg P. Cell surface trafficking of Fas: a rapid mechanism of p53-mediated apoptosis. Science 1998;282:290-3 https://doi.org/10.1126/science.282.5387.290
  23. Müller M, Wilder S, Bannasch D, Israeli D, Lehlbach K, Li-Weber M, et al. p53 activates the CD95 (APO-1/Fas) gene in response to DNA damage by anticancer drugs. J Exp Med 1998;188:2033-45 https://doi.org/10.1084/jem.188.11.2033
  24. Sheikh MS, Burns TF, Huang Y, Wu GS, Amundson S, Brooks KS, et al. p53-dependent and -independent regulation of the death receptor KILLER/DR5 gene expression in response to genotoxic stress and tumor necrosis factor alpha. Cancer Res 1988;58:1593-8
  25. Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, et al. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997;91:479-89 https://doi.org/10.1016/S0092-8674(00)80434-1
  26. Saleh A, Srinivasula SM, Acharya S, Fishel R, Alnemri ES. Cytochrome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation. J Biol Chem 1999;274:17941-5 https://doi.org/10.1074/jbc.274.25.17941
  27. Hu Y, Benedict MA, Ding L, Nunez G. Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis. EMBO J 1999;18:3586-95 https://doi.org/10.1093/emboj/18.13.3586
  28. Fuchs EJ, Mckenna KA, Bedi A. p53-dependent DNA damage-induced apoptosis requires Fas/APO-1-independent activation of CPP32beta. Cancer Res 1997;57:2550-4
  29. Henkels KM, Turchi JJ. Cisplatin-induced apoptosis proceeds by caspase-3-dependent and -independent pathways in cisplatin-resistant and -sensitive human ovarian cancer cell lines. Cancer Res 1999;59:3077-83
  30. Soengas MS, Alarcon RM, Yoshida H, Giaccia AJ, Hakem R, Mak TW, et al. Apaf-1 and caspase-9 in p53-dependent apoptosis and tumor inhibition. Science 1999;284:156-9 https://doi.org/10.1126/science.284.5411.156