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. 1999 May;80(5-6):693–698. doi: 10.1038/sj.bjc.6690411

Ceramide triggers p53-dependent apoptosis in genetically defined fibrosarcoma tumour cells

M Pruschy 1, H Resch 1, Y-Q Shi 1, N Aalame 1, C Glanzmann 1, S Bodis 1
PMCID: PMC2362278  PMID: 10360645

Abstract

p53 mutations are among the most common genetic alterations in human cancer and are frequently described in intrinsic or acquired radio- and chemotherapy resistance. Radiation-induced cell kill is not only mediated by DNA damage but also by the activation of signal transduction cascades generated at the plasma membrane like the sphingomyelin pathway. We used genetically defined wild-type p53 or p53-deficient mouse fibrosarcoma cells to investigate the p53-dependence of tumour response upon activation of the sphingomyelin pathway. Treatment of the tumour cells with neutral sphingomyelinase drastically reduced the amount of wild-type p53 fibrosarcoma cell proliferation over 72 h in a clear dose–response (0.2–1.0 U ml−1 nSMase). Sphingomyelinase had no effect on cell proliferation in tumour cells lacking p53. Similarly, cell proliferation was abolished by C2-ceramide (5–20 μM) only in wild-type p53 cells. FACS-analysis revealed that C2-ceramide induced massive p53-dependent apoptosis (40–50% after 12–24 h) and cell cycle analysis showed a transient G1 arrest in p53-deficient tumour cells 12–24 h after C2-ceramide exposure. These results suggest that ceramide-induced apoptosis in tumour cells can be dependent on the status of p53 and imply that p53 is also important for stress-induced apoptotic signal transduction cascades generated at the plasma membrane. © 1999 Cancer Research Campaign

Keywords: p53, ceramide, sphingomyelin pathway, radiation therapy

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Selected References

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  1. Alesse E., Zazzeroni F., Angelucci A., Giannini G., Di Marcotullio L., Gulino A. The growth arrest and downregulation of c-myc transcription induced by ceramide are related events dependent on p21 induction, Rb underphosphorylation and E2F sequestering. Cell Death Differ. 1998 May;5(5):381–389. doi: 10.1038/sj.cdd.4400358. [DOI] [PubMed] [Google Scholar]
  2. Amarante-Mendes G. P., Naekyung Kim C., Liu L., Huang Y., Perkins C. L., Green D. R., Bhalla K. Bcr-Abl exerts its antiapoptotic effect against diverse apoptotic stimuli through blockage of mitochondrial release of cytochrome C and activation of caspase-3. Blood. 1998 Mar 1;91(5):1700–1705. [PubMed] [Google Scholar]
  3. Chinnaiyan A. M., Orth K., O'Rourke K., Duan H., Poirier G. G., Dixit V. M. Molecular ordering of the cell death pathway. Bcl-2 and Bcl-xL function upstream of the CED-3-like apoptotic proteases. J Biol Chem. 1996 Mar 1;271(9):4573–4576. doi: 10.1074/jbc.271.9.4573. [DOI] [PubMed] [Google Scholar]
  4. Chmura S. J., Nodzenski E., Beckett M. A., Kufe D. W., Quintans J., Weichselbaum R. R. Loss of ceramide production confers resistance to radiation-induced apoptosis. Cancer Res. 1997 Apr 1;57(7):1270–1275. [PubMed] [Google Scholar]
  5. Clarke A. R., Purdie C. A., Harrison D. J., Morris R. G., Bird C. C., Hooper M. L., Wyllie A. H. Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature. 1993 Apr 29;362(6423):849–852. doi: 10.1038/362849a0. [DOI] [PubMed] [Google Scholar]
  6. Fisher D. E. Apoptosis in cancer therapy: crossing the threshold. Cell. 1994 Aug 26;78(4):539–542. doi: 10.1016/0092-8674(94)90518-5. [DOI] [PubMed] [Google Scholar]
  7. Haimovitz-Friedman A., Kan C. C., Ehleiter D., Persaud R. S., McLoughlin M., Fuks Z., Kolesnick R. N. Ionizing radiation acts on cellular membranes to generate ceramide and initiate apoptosis. J Exp Med. 1994 Aug 1;180(2):525–535. doi: 10.1084/jem.180.2.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haimovitz-Friedman A., Kolesnick R. N., Fuks Z. Ceramide signaling in apoptosis. Br Med Bull. 1997;53(3):539–553. doi: 10.1093/oxfordjournals.bmb.a011629. [DOI] [PubMed] [Google Scholar]
  9. Hannun Y. A. Apoptosis and the dilemma of cancer chemotherapy. Blood. 1997 Mar 15;89(6):1845–1853. [PubMed] [Google Scholar]
  10. Hannun Y. A., Linardic C. M. Sphingolipid breakdown products: anti-proliferative and tumor-suppressor lipids. Biochim Biophys Acta. 1993 Dec 21;1154(3-4):223–236. doi: 10.1016/0304-4157(93)90001-5. [DOI] [PubMed] [Google Scholar]
  11. Harris C. C. Structure and function of the p53 tumor suppressor gene: clues for rational cancer therapeutic strategies. J Natl Cancer Inst. 1996 Oct 16;88(20):1442–1455. doi: 10.1093/jnci/88.20.1442. [DOI] [PubMed] [Google Scholar]
  12. Herr I., Wilhelm D., Böhler T., Angel P., Debatin K. M. Activation of CD95 (APO-1/Fas) signaling by ceramide mediates cancer therapy-induced apoptosis. EMBO J. 1997 Oct 15;16(20):6200–6208. doi: 10.1093/emboj/16.20.6200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jacks T., Remington L., Williams B. O., Schmitt E. M., Halachmi S., Bronson R. T., Weinberg R. A. Tumor spectrum analysis in p53-mutant mice. Curr Biol. 1994 Jan 1;4(1):1–7. doi: 10.1016/s0960-9822(00)00002-6. [DOI] [PubMed] [Google Scholar]
  14. Jarvis W. D., Grant S., Kolesnick R. N. Ceramide and the induction of apoptosis. Clin Cancer Res. 1996 Jan;2(1):1–6. [PubMed] [Google Scholar]
  15. Jarvis W. D., Kolesnick R. N., Fornari F. A., Traylor R. S., Gewirtz D. A., Grant S. Induction of apoptotic DNA damage and cell death by activation of the sphingomyelin pathway. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):73–77. doi: 10.1073/pnas.91.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jayadev S., Liu B., Bielawska A. E., Lee J. Y., Nazaire F., Pushkareva MYu, Obeid L. M., Hannun Y. A. Role for ceramide in cell cycle arrest. J Biol Chem. 1995 Feb 3;270(5):2047–2052. doi: 10.1074/jbc.270.5.2047. [DOI] [PubMed] [Google Scholar]
  17. Kinscherf R., Claus R., Wagner M., Gehrke C., Kamencic H., Hou D., Nauen O., Schmiedt W., Kovacs G., Pill J. Apoptosis caused by oxidized LDL is manganese superoxide dismutase and p53 dependent. FASEB J. 1998 Apr;12(6):461–467. doi: 10.1096/fasebj.12.6.461. [DOI] [PubMed] [Google Scholar]
  18. Lowe S. W., Bodis S., McClatchey A., Remington L., Ruley H. E., Fisher D. E., Housman D. E., Jacks T. p53 status and the efficacy of cancer therapy in vivo. Science. 1994 Nov 4;266(5186):807–810. doi: 10.1126/science.7973635. [DOI] [PubMed] [Google Scholar]
  19. Lowe S. W., Jacks T., Housman D. E., Ruley H. E. Abrogation of oncogene-associated apoptosis allows transformation of p53-deficient cells. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2026–2030. doi: 10.1073/pnas.91.6.2026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lowe S. W., Ruley H. E., Jacks T., Housman D. E. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 1993 Sep 24;74(6):957–967. doi: 10.1016/0092-8674(93)90719-7. [DOI] [PubMed] [Google Scholar]
  21. Lowe S. W., Schmitt E. M., Smith S. W., Osborne B. A., Jacks T. p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature. 1993 Apr 29;362(6423):847–849. doi: 10.1038/362847a0. [DOI] [PubMed] [Google Scholar]
  22. Maity A., Kao G. D., Muschel R. J., McKenna W. G. Potential molecular targets for manipulating the radiation response. Int J Radiat Oncol Biol Phys. 1997 Feb 1;37(3):639–653. doi: 10.1016/s0360-3016(96)00598-6. [DOI] [PubMed] [Google Scholar]
  23. Miyashita T., Reed J. C. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell. 1995 Jan 27;80(2):293–299. doi: 10.1016/0092-8674(95)90412-3. [DOI] [PubMed] [Google Scholar]
  24. Obeid L. M., Linardic C. M., Karolak L. A., Hannun Y. A. Programmed cell death induced by ceramide. Science. 1993 Mar 19;259(5102):1769–1771. doi: 10.1126/science.8456305. [DOI] [PubMed] [Google Scholar]
  25. Santana P., Peña L. A., Haimovitz-Friedman A., Martin S., Green D., McLoughlin M., Cordon-Cardo C., Schuchman E. H., Fuks Z., Kolesnick R. Acid sphingomyelinase-deficient human lymphoblasts and mice are defective in radiation-induced apoptosis. Cell. 1996 Jul 26;86(2):189–199. doi: 10.1016/s0092-8674(00)80091-4. [DOI] [PubMed] [Google Scholar]
  26. Tsuchida E., Urano M. The effect of UCN-01 (7-hydroxystaurosporine), a potent inhibitor of protein kinase C, on fractionated radiotherapy or daily chemotherapy of a murine fibrosarcoma. Int J Radiat Oncol Biol Phys. 1997 Dec 1;39(5):1153–1161. doi: 10.1016/s0360-3016(97)00549-x. [DOI] [PubMed] [Google Scholar]
  27. Verheij M., Bose R., Lin X. H., Yao B., Jarvis W. D., Grant S., Birrer M. J., Szabo E., Zon L. I., Kyriakis J. M. Requirement for ceramide-initiated SAPK/JNK signalling in stress-induced apoptosis. Nature. 1996 Mar 7;380(6569):75–79. doi: 10.1038/380075a0. [DOI] [PubMed] [Google Scholar]
  28. Wyllie A. H. Apoptosis (the 1992 Frank Rose Memorial Lecture). Br J Cancer. 1993 Feb;67(2):205–208. doi: 10.1038/bjc.1993.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zhou H., Summers S. A., Birnbaum M. J., Pittman R. N. Inhibition of Akt kinase by cell-permeable ceramide and its implications for ceramide-induced apoptosis. J Biol Chem. 1998 Jun 26;273(26):16568–16575. doi: 10.1074/jbc.273.26.16568. [DOI] [PubMed] [Google Scholar]
  30. Zundel W., Giaccia A. Inhibition of the anti-apoptotic PI(3)K/Akt/Bad pathway by stress. Genes Dev. 1998 Jul 1;12(13):1941–1946. doi: 10.1101/gad.12.13.1941. [DOI] [PMC free article] [PubMed] [Google Scholar]

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