Apoptotic Response of Human Oral Squamous Carcinoma Cells to Etoposide

Etoposide에 대한 사람구강편평상피암종세포의 세포자멸사 반응

  • Kim, Gyoo-Cheon (Dept. of Oral Anatomy, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Lee, Kyoung-Duk (Dept. of Oral Anatomy, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Park, Jae-Hyun (Dept. of Oral Anatomy, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Kim, Duk-Han (Dept. of Oral Anatomy, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Park, Jeong-Kil (Dept. of Oral Conserve dentistry, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Park, June-Sang (Dept. of Oral Medicine, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Park, Bong-Soo (Dept. of Oral Anatomy, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University)
  • 김규천 (부산대학교 치과대학 및 구강생물공학연구소 구강해부학교실) ;
  • 이경덕 (부산대학교 치과대학 및 구강생물공학연구소 구강해부학교실) ;
  • 박재현 (부산대학교 치과대학 및 구강생물공학연구소 구강해부학교실) ;
  • 김덕한 (부산대학교 치과대학 및 구강생물공학연구소 구강해부학교실) ;
  • 박정길 (부산대학교 치과대학 및 구강생물공학연구소 구강보존과학교실) ;
  • 박준상 (부산대학교 치과대학 및 구강생물공학연구소 구강내과학교실) ;
  • 박봉수 (부산대학교 치과대학 및 구강생물공학연구소 구강해부학교실)
  • Published : 2005.06.30


Anti-cancer drugs have been shown to target diverse cellular functions in mediation cell death in chemosensitive tumors. Most antineoplastic drugs used in chemotherapy of leukemias and solid tumors induce apoptosis in drug-sensitive target cells. However, the precise molecular requirements that are central for drug-induced cell death are largely unknown. Etoposide is used for the treatment of lung and testicular cancer. This study was performed to examine whether etoposide promote apoptosis in human oral squamous carcinoma cells (OSC9) as well as in lung and testicular cancer. Etoposide had a significant dose- and time-dependent inhibitory effect on the viability of OSC9 cells. TUNEL assay showed the positive reaction on condensed nuclei. Hoechst stain demonstrated that etoposide induced a change in nuclear morphology. The expression of p53 was increased at 48 hour, suggesting that the nuclear of OSC9 cell was damaged, thereby inducing apoptosis. Etoposide treatment induced caspase-3 cleavage and activation. Intact PARP protein 116-kDa and 85-kDa cleaved product were observed. The activated caspase-3 led cleavage of the PARP. These results demonstrate that etoposide-induced apoptosis in OSC9 cells is associated with caspase-3 activation.


  1. Wyllie, A.H., Kerr, J.F., and Currie, A.R.: Cell death: the significance of apoptosis. Int Rev Cytol 68 :251-306, 1980 https://doi.org/10.1016/S0074-7696(08)62312-8
  2. Golstein P: Controlling cell death. Science 275(5303) :1081-1082, 1997 https://doi.org/10.1126/science.275.5303.1081
  3. Orth, K., O'Rourke, K., Salvesen, G.S., and Dixit, V.M.: Molecular ordering of apoptotic mammalian CED-3/ICE-like proteases. J Biol Chem 271(35) :20977-20980, 1996 https://doi.org/10.1074/jbc.271.35.20977
  4. Enari, M., Sakahira, H., Yokoyama, H. et al. : A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391(6662) :43-50, 1998 https://doi.org/10.1038/34112
  5. Sakahira, H., Enari, M. and Nagata, S.: Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391(6662) :96-99, 1998 https://doi.org/10.1038/34214
  6. Thornberry, N.A., Rosen, A. and Nicholson, D.W.: Control of apoptosis by proteases, in Apoptosis (Kaufmann SH eds). Adv Pharmacol 44 : 155-177, 1997
  7. Yuan, J.: Evolutionary conservation of a genetic pathway of programmed cell death. J Cell Biochem 60: 4-11, 1996 https://doi.org/10.1002/(SICI)1097-4644(19960101)60:1<4::AID-JCB2>3.0.CO;2-1
  8. Clarke, A.R., Purdie, C.A., Harrison, D.J. et al. : Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 362(6423) : 849-852, 1993 https://doi.org/10.1038/362849a0
  9. Wagner, A.J., Kokontis, J.M. and Hay, N.: Myc-mediated apoptosis requires wild-type p53 in a manner independent of cell cycle arrest and the ability of p53 to induce p21waf1/cip1. Genes Dev 8 :2817-2830, 1994 https://doi.org/10.1101/gad.8.23.2817
  10. Graeber, T.G., Osmanian, C., Jacks, T., Housman, D.E., Koch, C.J., Lowe, S.W. and Giaccia, A.J.: Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours. Nature 379 :88-91, 1996 https://doi.org/10.1038/379088a0
  11. Linke, S.P., Clarkin, K.C., Di, Leonardo, A., Tsou, A. and Wahl, G.M.: A reversible, p53-dependent G0/G1 cell cycle arrest induced by ribonucleotide depletion in the absence of detectable DNA damage. Genes Dev 10 : 934-947, 1996 https://doi.org/10.1101/gad.10.8.934
  12. Yin, Y., Terauchi, Y., Solomon, G.G. et al. : Involvement of p85 in p53-dependent apoptotic response to oxidative stress. Nature 391 : 707-710, 1998 https://doi.org/10.1038/35648
  13. Schmitt, C.A. and Lowe, S.W. : Apoptosis and Therapy. J Pathol 187 : 127-137, 1999 https://doi.org/10.1002/(SICI)1096-9896(199901)187:1<127::AID-PATH251>3.0.CO;2-T
  14. Attardi, L.D., Reczek, E.E., Cosmas, C. et al. : PERP, an apoptosis-associated target of p53, is a novel member of the PMP-22/gas3 family. Genes Dev 14 : 704-718, 2000
  15. Ko, L.J. and Prives, C.: p53: puzzle and paradigm. Genes Dev 10 : 1054-1072, 1996 https://doi.org/10.1101/gad.10.9.1054
  16. Levine, A.J.: The tumor suppressor genes. Annu Rev Biochem 62 : 623-651, 1993 https://doi.org/10.1146/annurev.bi.62.070193.003203
  17. Fisher, D.E.: Apoptosis in cancer therapy: crossing the threshold. Cell 78(4) : 539-542, 1994 https://doi.org/10.1016/0092-8674(94)90518-5
  18. Steller, H.: Mechanisms and genes of cellular suicide. Science 267(5203) : 1445-1449, 1995 https://doi.org/10.1126/science.7878463
  19. Lowe, S.W., Ruley, H.E., Jacks, T. and Housman, D.E.: p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74(6) : 957-967, 1993 https://doi.org/10.1016/0092-8674(93)90719-7
  20. Burden, D.A., Kingma, P.S., Froelich-Ammon, S.J. et al. : Topoisomerase II.etoposide interactions direct the formation of drug-induced enzyme-DNA cleavage complexes. J Biol Chem 271(46) : 29238-29244, 1996 https://doi.org/10.1074/jbc.271.46.29238
  21. Ramirez, C.D., Sleiman, R.J., Catchpoole, D.R. and Stewart, B.W.: Morphological and molecular evidence of differentiation during etoposide-induced apoptosis in human lymphoblastoid cells. Cell Death Differ 7(6): 548-555, 2000 https://doi.org/10.1038/sj.cdd.4400686
  22. Ucker, D.S., Obermiller, P.S., Eckhart, W. et al. : Genome digestion is a dispensable consequence of physiological cell death mediated by cytotoxic T lymphocytes. Mol Cell Biol 12 : 3060-3069, 1992 https://doi.org/10.1128/MCB.12.7.3060
  23. Nicholson, D.W. and Thornberry, N.A.: Caspases: killer proteases. Trends Biochem Sci 22 : 299-306, 1997 https://doi.org/10.1016/S0968-0004(97)01085-2
  24. Schulze-Osthoff, K., Walczak, H., Droge, W. and Krammer, P.H.: Cell nucleus and DNA fragmentation are not required for apoptosis. J Cell Biol 127 : 15-20, 1994 https://doi.org/10.1083/jcb.127.1.15
  25. Williams, G.T. : Programmed cell death: apoptosis and oncogenesis. Cell 65 : 1097-1098, 1991 https://doi.org/10.1016/0092-8674(91)90002-G
  26. Tolis, C., Peters, G.J., Ferreira, C.G., Pinedo, H.M. and Giaccone, G.: Cell cycle disturbances and apoptosis induced by topotecan and gemcitabine on human lung cancer cell lines. Eur J Cancer 35(5) : 796-807, 1999 https://doi.org/10.1016/S0959-8049(98)00425-0
  27. Harris, C.C.: Structure and function of the p53 tumor suppressor gene: clues for rational cancer therapeutic strategies. J Natl Cancer Inst 88(20) : 1442-1455, 1996 https://doi.org/10.1093/jnci/88.20.1442
  28. Soldani, C., Lazze, M.C., Bottone, M.G. et al. : Poly(ADP-ribose) polymerase cleavage during apoptosis: when and where? Exp Cell Res 269(2) :193-201, 2001 https://doi.org/10.1006/excr.2001.5293