Synthetic Chenodeoxycholic Acid Derivative HS-1200-Induced Apoptosis of Human Oral Squamous Carcinoma Cells

합성 Chenodeoxycholic Acid 유도체 HS-1200이 유도한 사람구강 편평상피암종세포 세포자멸사 연구

  • Kim, In-Ryoung (Department of Biology, College of Natural Science, Pusan National University) ;
  • Sohn, Hyeon-Jin (Department of Oral Anatomy, College of Dentistry, Pusan National University) ;
  • Kim, Gyoo-Cheon (Department of Oral Anatomy, College of Dentistry, Pusan National University) ;
  • Kwak, Hyun-Ho (Department of Oral Anatomy, College of Dentistry, Pusan National University) ;
  • Park, Bong-Soo (Department of Oral Anatomy, College of Dentistry, Pusan National University) ;
  • Choi, Won-Chul (Department of Biology, College of Natural Science, Pusan National University) ;
  • Ko, Myung-Yun (Department of Oral Medicine, College of Dentistry, Pusan National University) ;
  • Ahn, Yong-Woo (Department of Oral Medicine, College of Dentistry, Pusan National University)
  • 김인령 (부산대학교 자연과학대학 생물학과) ;
  • 손현진 (부산대학교 치과대학 구강해부학교실) ;
  • 곽현호 (부산대학교 치과대학 구강해부학교실) ;
  • 김규천 (부산대학교 치과대학 구강해부학교실) ;
  • 박봉수 (부산대학교 치과대학 구강해부학교실) ;
  • 최원철 (부산대학교 자연과학대학 생물학과) ;
  • 고명연 (부산대학교 치과대학 구강내과학교실) ;
  • 안용우 (부산대학교 치과대학 구강내과학교실)
  • Published : 2007.09.30


Bile acids and synthetic its derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. Previous studies have been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis inducing activity on various cancer cells in vitro. It wasn't discovered those materials have apoptosis induced effects on YD9 human oral squamous carcinoma cells. The present study was done to examine the synthetic bile acid derivatives(HS-1199, HS-1200) induced apoptosis on YD9 cells and such these apoptosis events. We administered them in culture to YD9 cells. Tested YD9 cells showed several lines of apoptotic manifestation such as activation of caspase-3, degradation of DFF, production of poly (ADP-ribose) polymerase(PARP) cleavage(HS-1200 only), DNA degradation(HS-1200 only), nuclear condensation, inhibition of proteasome activity, reduction of mitochondrial membrane potential(HS-1200 only) and the release of cytochrome c and AIF to cytosol. Between two synthetic CDCA derivatives, HS-1200 showed stronger apoptosis-inducing effect than HS-1199. Therefore HS-1200 was demonstrated to have the most efficient antitumor effect. Taken collectively, we demonstrated that a synthetic CDCA derivative HS-1200 induced caspases-dependent apoptosis via mitochondrial pathway in human oral sqauamous carcinoma cells in vitro. Our data therefore provide the possibility that HS-1200 could be considered as a novel therapeutic strategy for human orall squamous carcinoma from its poweful apoptosis-inducing activity.

담즙산과 합성담즙산유도체가 여러 종류의 암세포에 세포자멸사(apoptosis)를 유도하고 항암효과가 있다고 알려져 있다. 합성 chenodeoxycholic acid (CDCA) 유도체가 여러 가지 암세포에 유도한 세포자멸사 in vitro 연구들이 보고되어져 왔다. 하지만 아직 까지 구강편평상피암종세포에 합성 CDCA 유도체가 유도한 세포자멸사 연구는 없었다. 그래서 본 연구는 합성 CDCA 유도체인 HS-1199와 HS-1200이 사람구강편평상피암종세포에 세포자멸사 효과와 세포자멸사 기작을 알기 위해서 수행되었다. 합성 CDCA 유도체로 처리된 사람구강편평상피암종세포(YD9 세포)에서 caspase-3의 활성화, DFF의 degradation, poly (ADP-ribose) polymerase(PARP)의 분절화(HS-1200 only), DNA 분절화(HS-1200 only), 핵 응축, proteosome 활성화의 저해, 사립체막전위 (MMP)의 감소(HS-1200 only) 그리고 cytochrome c와 AIF의 사립체에서 세포질로의 유리와 같은 세포자멸사의 증거를 보였다. 그리고 두 개의 합성 CDCA 유도체 중에서 HS-1200이 HS-1199보다 더욱 더 강한 세포자멸사 효과를 보였다. 이 결과는 HS-1200이 YD9 세포에 항암효과를 가진다는 것을 증명한 것이다. 본 연구는 CDCA 유도체인 HS-1200이 사람구강편평상피암종세포에서 사립체 경로를 통한 caspase 의존적 세포자멸사를 강력하게 유도한다는 것을 증명했으며, 이러한 결과는 HS-1200이 사람구강편평상피암종의 치료적 전략으로서의 가능성이 높다고 생각한다.



  1. Shen J, Huang C, Jiang L et al.. Enhancement of cisplatin induced apoptosis by suberoylanilide hydroxamic acid in human oral squamous cell carcinoma cell lines. Biochem Pharmacol 2007;73:1901-1909
  2. Zimmermann KC, Bonzon C, Green DR. The machinery of programmed cell death. Pharmacol Ther 2001;92:57-70
  3. Holtzman MJ, Green JM, Jayaraman S, Arch RH. Regulation of T cell apoptosis. Apoptosis 2000;5:459-471
  4. Reed JC. Apoptosis-regulating proteins as targets for drug discovery. Trend Mol Med 2001;5:459-471
  5. Kankaanranta H, Giembycz MA, Barnes PJ, el- Haddad B, Saarelainen S, Zhang X, Moilanen E, Lindsay MA. Hydrogen peroxide reverses IL-5 afforded eosinophil survival and promotes constitutive human eosinophil apoptosis. Int Arch Allergy Immunol 2002;127:73-78
  6. Hofmann AF. Chemistry and enterohepatic circulation of bile acids. Hepatology 1984;4(5 Suppl):4S-14S
  7. Martinez JD, Stratagoules ED, LaRue JM et al. Different bile acids exhibit distinct biological effects: the tumor promoter deoxycholic acid induces apoptosis and the chemopreventive agent ursodeoxycholic acid inhibits cell proliferation. Nutr Cancer 1998;31:111-118
  8. Jones B, Roberts PJ, Faubion WA, Kominami E, Gores GJ. Cystatin A expression reduces bile salt-induced apoptosis in a rat hepatoma cell line. Am J Physiol 1998;275:723-730
  9. Martinez-Diez MC, Serrano MA, Monte MJ, Marin JJ. Comparison of the effects of bile acids on cell viability and DNA synthesis by rat hepatocytes in primary culture. Biochim Biophys Acta 2000;1500: 153-160
  10. Rust C, Karnitz LM, Paya CV, Moscat J, Simari RD, Gores GJ. The bile acid taurochenodeoxycholate activates a phosphatidylinositol 3-kinase-dependent survival signaling cascade. J Biol Chem 2000;275: 20210-20216
  11. Patel T, Bronk SF, Gores GJ. Increases of intracellular magnesium promote glycodeoxycholate-induced apoptosis in rat hepatocytes. J Clin Invest 1994;94:2183-2192
  12. Patel T, Roberts LR, Jones BA, Gores GJ. Dysregulation of apoptosis as a mechanism of liver disease: an overview. Semin Liver Dis 1998;18:105-114
  13. Powell AA, LaRue JM, Batta AK, Martinez JD. Bile acid hydrophobicity is correlated with induction of apoptosis and/or growth arrest in HCT116 cells. Biochem J 2001;356:481-486
  14. Im EO, Choi YH, Paik KJ et al. Novel bile acid derivatives induce apoptosis via a p53-independent pathway in human breast carcinoma cells. Cancer Lett 2001;163:83-93
  15. Wyllie AH, Kerr JFR, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol 1980;68:251-305
  16. Williams GT. Programmed cell death: apoptosis and oncogenesis. Cell 1991;65:1097-1098
  17. Thornberry NA, Rosen A, Nicholson DW. Control of apoptosis by proteases, in Apoptosis (Kaufmann SH eds). Adv Pharmacol 1997;44:155-177
  18. Yuan J. Evolutionary conservation of a genetic pathway of programmed cell death. J Cell Biochem 1996;60:4-11<4::AID-JCB2>3.0.CO;2-1
  19. Choi Y H, Im EO, Suh H et al. Apoptotic activity of novel bile acid derivatives in human leukemic T cells through the activation of caspases. Int J Oncol 2001;18:979-984
  20. Seo SY, Jun EJ, Jung SM et al. Synthetic chenodeoxycholic acid derivative HS-1200-induced apoptosis of p815 mastocytoma cells is augmented by co-treatment with lactacystin. Anticancer Drugs 2003;14:219-225
  21. Jeong JH, Park JS, Moon B et al. Orphan nuclear receptor Nur77 translocates to mitochondria in the early phase of apoptosis induced by synthetic chenodeoxycholic acid derivatives in human stomach cancer cell line SNU-1. Ann N Y Acad Sci 2003;1010:171-177
  22. Choi HJ, Kim HH, Lee HS et al. Lactacystin augments the sulindac-induced apoptosis in HT-29 cells. Apoptosis 2003;8:301-305
  23. Yoon HS, Rho JH, Yoo KW et al. Synthetic bile acid derivatives induce nonapoptotic death of human retinal pigment epithelial cells. Curr Eye Res 2001;22:367-374
  24. Drexler HC, Risau W, Konerding MA. Inhibition of proteasome function induces programmed cell death in proliferating endothelial cells. FASEB J 2000;14: 65-77
  25. Orlowski RZ. The role of the ubiquitin-proteasome pathway in apoptosis. Cell Death Differ 1999;6: 303-313
  26. Grimm LM, Goldberg AL, Poirier GG, Schwartz LM, Osborne BA. Proteasomes play an essential role in thymocyte apoptosis. EMBO J 1996;15:3835-3844
  27. Sadoul R, Fernandez PA, Quiquerez AL et al. Involvement of the proteasome in the programmed cell death of NGF-deprived sympathetic neurons. EMBO J 1996;15:3845-3852
  28. Li B, Dou QP. Bax degradation by the ubiquitin/ proteasome-dependent pathway: involvement in tumor survival and progression. Proc Natl Acad Sci USA 2000;97:3850-3855
  29. Orlowski RZ, Eswara JR, Lafond-Walker A, Grever MR, Orlowski M, Dang CV. Tumor growth inhibition induced in a murine model of human Burkitt's lymphoma by a proteasome inhibitor. Cancer Res 1998;58:4342-4348
  30. Adams J, Palombella VJ, Sausville EA et al. Proteasome inhibitors: a novel class of potent and effective antitumor agents. Cancer Res 1999;59:2615-2622
  31. Chandra J, Niemer I, Gilbreath J et al. Proteasome inhibitors induce apoptosis in glucocorticoid-resistant chronic lymphocytic leukemic lymphocytes. Blood 1998;92: 4220-4229
  32. Delic J, Masdehors P, Omura S et al. The proteasome inhibitor lactacystin induces apoptosis and sensitizes chemo- and radioresistant human chronic lymphocytic leukaemia lymphocytes to TNF-alphainitiated apoptosis. Br J Cancer 1998;77:1103-1107
  33. Fanelli M, Minucci S, Gelmetti V et al. Constitutive degradation of PML/RARalpha through the proteasome pathway mediates retinoic acid resistance. Blood 1999;93:1477-1481
  34. Golab J, Stoklosa T, Czajka A et al. Synergistic antitumor effects of a selective proteasome inhibitor and TNF in mice. Anticancer Res 2000;20:1717-1721
  35. Park BS, Baek SJ, Song KH et al. Genistein-induced apoptosis of p815 mastocytoma cells is mediated by Bax and augmented by a proteasome inhibitor, lactacystin. Nutr Cancer 2002;42:248-255
  36. Kroemer G, Zamzami N, Susin SA. Mitochondrial control of apoptosis. Immunol Today 1997;18:44-51
  37. Green DR, Reed JC. Mitochondria and apoptosis. Science 1998;281:1309-1312
  38. Susin SA, Lorenzo HK, Zamzami N et al. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 1999;397:441-446
  39. Orrenius S. Mitochondrial regulation of apoptotic cell death. Toxicol. Lett 2004;149:19-23
  40. Li P, Nijhawan D, Budihardjo I et al. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997;91:479-489
  41. Wagenknecht B, Hermisson M, Groscurth P, Liston P, Krammer PH, Weller M. Proteasome inhibitorinduced apoptosis of glioma cells involves the processing of multiple caspases and cytochrome c release. J Neurochem 2000;75:2288-2297
  42. Marshansky V, Wang X, Bertrand R et al. Proteasomes modulate balance among proapoptotic and antiapoptotic Bcl-2 family members and compromise functioning of the electron transport chain in leukemic cells. J Immunol 2001;166:3130-3142
  43. Daugas E, Susin SA, Zamzami N et al. Mitochondrio -nuclear translocation of AIF in apoptosis and necrosis. FASEB J 2000;14:729-739
  44. Liu X, Li P, Widlak P et al. The 40-kDa subunit of DNA fragmentation factor induces DNA fragmentation and chromatin condensation during apoptosis. Proc Natl Acad Sci USA 1998;95:8461-8466