Apoptotic Effect of co-treatment with HS-1200 and Cisplatin on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line

HS-1200과 cisplatin의 병용처리가 사람구강암세포에 미치는 세포자멸사 효과에 대한 연구

  • Kim, Duk-Han (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kim, In-Ryoung (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Park, Bong-Soo (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Ahn, Yong-Woo (Department of Oral Medicine, School of Dentistry, Pusan National University) ;
  • Jeong, Sung-Hee (Department of Oral Medicine, School of Dentistry, Pusan National University)
  • 김덕한 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 김인령 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 박봉수 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 안용우 (부산대학교 치의학전문대학원 구강내과학교실) ;
  • 정성희 (부산대학교 치의학전문대학원 구강내과학교실)
  • Received : 2013.06.18
  • Accepted : 2013.08.22
  • Published : 2013.09.30


Bile acids are polar derivatives of cholesterol essential for the absorption of dietary lipids and regulate the transcription of genes that control cholesterol homeostasis. Recently it have been identified the synthetic chenodeoxycholic acid (CDCA) derivatives HS-1200 and cisplatin showed apoptisis-inducing activity on various cancer cells in vivo and in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with HS-1200 and cisplatin on human tongue squamous cell carcinoma cells (SCC25 cells). To investigate whether the co-treatment with HS-1200 and cisplatin compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis DNA hypoploidy. Westen blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. In this study, co-treatment with HS-1200 and cisplatin on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, reduction of MMP and proteasome activity, the increase of Bax and the decrease of Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD) whereas each single treated SCC25 cells did not show these patterns. Although the single treatment of $25{\mu}M$ HS-1200 and $4{\mu}g/ml$ cisplatin for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that the combination therapy with HS-1200 and cisplatin could be considered as a novel therapeutic strategy for human squamous cell carcinoma.


Supported by : Pusan National University


  1. Williams GT. Programmed cell death: apoptosis and oncogenesis. Cell 1991;65:1097-1098.
  2. 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
  3. Susin SA, Lorenzo HK, Zamzami N et al. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 1999;397:441-446.
  4. Orlowski RZ. The role of the ubiquitin-proteasome pathway in apoptosis. Cell Death Differ 1999;6: 303-313.
  5. 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.
  6. 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:G723-730.
  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. 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.
  9. Im EO, Lee S, Suh H, Kim KW, Bae YT, Kim ND. A novel ursodeoxycholic acid derivative induces apoptosis in human MCF-7 breast cancer cells. Pharm Pharmacol Commun 1999;5:293-298.
  10. 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.
  11. 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.
  12. Park SE, Choi HJ, Yee SB et al. Synthetic bile acid derivatives inhibit cell proliferation and induce apoptosis in HT-29 human colon cancer cells. Int J Oncol 2004;25:231-236.
  13. Im E, Choi SH, Suh H et al. Synthetic bile acid derivatives induce apoptosis through a c-Jun N-terminal kinase and NF-kappaB-dependent process in human cervical carcinoma cells. Cancer Lett 2005;229:49-57.
  14. Kim ND, Im E, Yoo YH, Choi YH. Modulation of the cell cycle and induction of apoptosis in human cancer cells by synthetic bile acids. Curr Cancer Drug Targets 2006;6:681-689.
  15. Wang G, Reed E, Li QQ. Molecular basis of cellular response to cisplatin chemotherapy in non-small cell lung cancer (Review). Oncol Rep 2004;12: 955-965.
  16. Choi YH, 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.
  17. 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.
  18. Choi YH, Im EO, Suh H et al. Apoptosis and modulation of cell cycle control by synthetic derivatives of ursodeoxycholic acid and chenodeoxycholic acid in human prostate cancer cells. Cancer Lett 2003;199:157-167.
  19. Cooley ME, Davis LE, DeStefano MAbrahm J. Cisplatin: a clinical review. Part I--Current uses of cisplatin and administration guidelines. Cancer Nurs 1994;17:173-184.
  20. Gonzalez VM, Fuertes MA, Alonso C, Perez JM. Is cisplatin-induced cell death always produced by apoptosis? Mol Pharmacol 2001;59:657-663.
  21. Seki K, Yoshikawa H, Shiiki K et al. Cisplatin (CDDP) specifically induces apoptosis via sequential activation of caspase-8, -3 and -6 in osteosarcoma. Cancer Chemother Pharmacol 2000;45:199-206.
  22. Del Bello B, Valentini MA, Comporti M, Maellaro E. Cisplatin-induced apoptosis in melanoma cells: role of caspase-3 and caspase-7 in Apaf-1 proteolytic cleavage and in execution of the degradative phases. Ann N Y Acad Sci 2003;1010:200-204.
  23. Zhou H, Kato A, Yasuda H et al. The induction of cell cycle regulatory and DNA repair proteins in cisplatin-induced acute renal failure. Toxicol Appl Pharmacol 2004;200:111-120.
  24. Fox SA, Kusmiaty, Loh SS, Dharmarajan AM, Garlepp MJ. Cisplatin and TNF-alpha downregulate transcription of Bcl-xL in murine malignant mesothelioma cells. Biochem Biophys Res Commun 2005;337:983-991.
  25. Yde CW, Issinger OG. Enhancing cisplatin sensitivity in MCF-7 human breast cancer cells by down-regulation of Bcl-2 and cyclin D1. Int J Oncol 2006;29:1397-1404.
  26. Garcia-Berrocal JR, Nevado J, Ramirez-Camacho R et al. The anticancer drug cisplatin induces an intrinsic apoptotic pathway inside the inner ear. Br J Pharmacol 2007;152:1012-1020.
  27. Gagnon V, Van Themsche C, Turner S, Leblanc V, Asselin E. Akt and XIAP regulate the sensitivity of human uterine cancer cells to cisplatin, doxorubicin and taxol. Apoptosis 2008;13:259-271.
  28. Kondo K, Yamasaki S, Inoue N et al. Prospective antitumor effects of the combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and cisplatin against esophageal squamous cell carcinoma. Surg Today 2006;36:966-974.
  29. Mohammad RM, Banerjee S, Li Y et al. Cisplatin-induced antitumor activity is potentiated by the soy isoflavone genistein in BxPC-3 pancreatic tumor xenografts. Cancer 2006;106:1260-1268.
  30. Iwase M, Yoshiba S, Uchid M et al. Enhanced susceptibility to apoptosis of oral squamous cell carcinoma cells subjected to combined treatment with anticancer drugs and phosphatidylinositol 3-kinase inhibitors. Int J Oncol 2007;31:1141-1147.
  31. Pisano C, Vesci L, Fodera R et al. Antitumor activity of the combination of synthetic retinoid ST1926 and cisplatin in ovarian carcinoma models. Ann Oncol 2007;18:1500-1505.
  32. Drexler HC, Risau W, Konerding MA. Inhibition of proteasome function induces programmed cell death in proliferating endothelial cells. FASEB J 2000;14: 65-77.
  33. Kroemer G. Mitochondrial control of apoptosis: an introduction. Biochem Biophys Res Commun 2003; 304:433-435.
  34. Green DR, Reed JC. Mitochondria and apoptosis. Science 1998;281:1309-1312.
  35. 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.
  36. Wagenknecht B, Hermisson M, Groscurth P et al. Proteasome inhibitor-induced apoptosis of glioma cells involves the processing of multiple caspases and cytochrome c release. J Neurochem 2000;75:2288- 2297.
  37. 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.
  38. Daugas E, Nochy D, Ravagnan L et al. Apoptosisinducing factor (AIF): a ubiquitous mitochondrial oxidoreductase involved in apoptosis. FEBS Lett 2000;476:118-123.
  39. Acehan D, Jiang X, Morgan DG et al. Threedimensional structure of the apoptosome: implications for assembly, procaspase-9 binding, and activation. Mol Cell 2002;9:423-432.
  40. Gross A, McDonnell JM, Korsmeyer SJ. BCL-2 family members and the mitochondria in apoptosis. Genes Dev 1999;13:1899-1911.
  41. Porter AG. Protein translocation in apoptosis. Trends Cell Biol 1999;9:394-401.
  42. Cheng AC, Jian CB, Huang YT et al. Induction of apoptosis by Uncaria tomentosa through reactive oxygen species production, cytochrome c release, and caspases activation in human leukemia cells. Food Chem Toxicol 2007;45:2206-2218.