Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2003 Oct 15;375(Pt 2):263–274. doi: 10.1042/BJ20030868

Bcl-2 activates a programme of premature senescence in human carcinoma cells.

Elvira Crescenzi 1, Giuseppe Palumbo 1, Hugh J M Brady 1
PMCID: PMC1223693  PMID: 12871207

Abstract

The apoptosis regulator Bcl-2 has been shown to modulate cell-cycle progression, favouring a quiescent state over a proliferative state, in both normal and tumour cells. We show here that constitutive expression of Bcl-2 in human carcinoma cells results in a cell-cycle arrest that within a few days can become irreversible. Arrested cells acquire a senescent-like phenotype, which consists of several characteristic morphological alterations and increased activity of senescence-associated beta-galactosidase. The induction of the premature senescence programme is mediated by inhibition of Cdk2 kinase activity, and p27(KIP1) is required to maintain the senescent phenotype. We propose that the ability to activate an endogenous premature senescence programme allows Bcl-2 to suppress tumour growth. These results suggest that the down-regulation of Bcl-2 expression, which has been observed during the development and progression of human carcinoma, is related to the ability of Bcl-2 to severely hamper the growth of carcinoma cells and to induce a permanent cell-cycle arrest, with the features of senescence.

Full Text

The Full Text of this article is available as a PDF (349.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alexander K., Hinds P. W. Requirement for p27(KIP1) in retinoblastoma protein-mediated senescence. Mol Cell Biol. 2001 Jun;21(11):3616–3631. doi: 10.1128/MCB.21.11.3616-3631.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biden K. G., Simms L. A., Cummings M., Buttenshaw R., Schoch E., Searle J., Gobe G., Jass J. R., Meltzer S. J., Leggett B. A. Expression of Bcl-2 protein is decreased in colorectal adenocarcinomas with microsatellite instability. Oncogene. 1999 Feb 4;18(5):1245–1249. doi: 10.1038/sj.onc.1202413. [DOI] [PubMed] [Google Scholar]
  3. Brady H. J., Gil-Gómez G., Kirberg J., Berns A. J. Bax alpha perturbs T cell development and affects cell cycle entry of T cells. EMBO J. 1996 Dec 16;15(24):6991–7001. [PMC free article] [PubMed] [Google Scholar]
  4. Bringold F., Serrano M. Tumor suppressors and oncogenes in cellular senescence. Exp Gerontol. 2000 May;35(3):317–329. doi: 10.1016/s0531-5565(00)00083-8. [DOI] [PubMed] [Google Scholar]
  5. Chang B. D., Broude E. V., Dokmanovic M., Zhu H., Ruth A., Xuan Y., Kandel E. S., Lausch E., Christov K., Roninson I. B. A senescence-like phenotype distinguishes tumor cells that undergo terminal proliferation arrest after exposure to anticancer agents. Cancer Res. 1999 Aug 1;59(15):3761–3767. [PubMed] [Google Scholar]
  6. Chen Y. N., Sharma S. K., Ramsey T. M., Jiang L., Martin M. S., Baker K., Adams P. D., Bair K. W., Kaelin W. G., Jr Selective killing of transformed cells by cyclin/cyclin-dependent kinase 2 antagonists. Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4325–4329. doi: 10.1073/pnas.96.8.4325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chestukhin Anton, Litovchick Larisa, Rudich Katherine, DeCaprio James A. Nucleocytoplasmic shuttling of p130/RBL2: novel regulatory mechanism. Mol Cell Biol. 2002 Jan;22(2):453–468. doi: 10.1128/MCB.22.2.453-468.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Collado M., Medema R. H., Garcia-Cao I., Dubuisson M. L., Barradas M., Glassford J., Rivas C., Burgering B. M., Serrano M., Lam E. W. Inhibition of the phosphoinositide 3-kinase pathway induces a senescence-like arrest mediated by p27Kip1. J Biol Chem. 2000 Jul 21;275(29):21960–21968. doi: 10.1074/jbc.M000759200. [DOI] [PubMed] [Google Scholar]
  9. Day M. L., Foster R. G., Day K. C., Zhao X., Humphrey P., Swanson P., Postigo A. A., Zhang S. H., Dean D. C. Cell anchorage regulates apoptosis through the retinoblastoma tumor suppressor/E2F pathway. J Biol Chem. 1997 Mar 28;272(13):8125–8128. doi: 10.1074/jbc.272.13.8125. [DOI] [PubMed] [Google Scholar]
  10. Dimri G. P., Lee X., Basile G., Acosta M., Scott G., Roskelley C., Medrano E. E., Linskens M., Rubelj I., Pereira-Smith O. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9363–9367. doi: 10.1073/pnas.92.20.9363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dulić V., Lees E., Reed S. I. Association of human cyclin E with a periodic G1-S phase protein kinase. Science. 1992 Sep 25;257(5078):1958–1961. doi: 10.1126/science.1329201. [DOI] [PubMed] [Google Scholar]
  12. Evan G. I., Vousden K. H. Proliferation, cell cycle and apoptosis in cancer. Nature. 2001 May 17;411(6835):342–348. doi: 10.1038/35077213. [DOI] [PubMed] [Google Scholar]
  13. Furth P. A., Bar-Peled U., Li M., Lewis A., Laucirica R., Jäger R., Weiher H., Russell R. G. Loss of anti-mitotic effects of Bcl-2 with retention of anti-apoptotic activity during tumor progression in a mouse model. Oncogene. 1999 Nov 11;18(47):6589–6596. doi: 10.1038/sj.onc.1203073. [DOI] [PubMed] [Google Scholar]
  14. Geisler J. P., Geisler H. E., Wiemann M. C., Zhou Z., Miller G. A., Crabtree W. Lack of bcl-2 persistence: an independent prognostic indicator of poor prognosis in endometrial carcinoma. Gynecol Oncol. 1998 Nov;71(2):305–307. doi: 10.1006/gyno.1998.5192. [DOI] [PubMed] [Google Scholar]
  15. Gil-Gómez G., Berns A., Brady H. J. A link between cell cycle and cell death: Bax and Bcl-2 modulate Cdk2 activation during thymocyte apoptosis. EMBO J. 1998 Dec 15;17(24):7209–7218. doi: 10.1093/emboj/17.24.7209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hanahan D., Weinberg R. A. The hallmarks of cancer. Cell. 2000 Jan 7;100(1):57–70. doi: 10.1016/s0092-8674(00)81683-9. [DOI] [PubMed] [Google Scholar]
  17. Hannon G. J., Demetrick D., Beach D. Isolation of the Rb-related p130 through its interaction with CDK2 and cyclins. Genes Dev. 1993 Dec;7(12A):2378–2391. doi: 10.1101/gad.7.12a.2378. [DOI] [PubMed] [Google Scholar]
  18. Kaufmann J. A., Bickford P. C., Taglialatela G. Oxidative-stress-dependent up-regulation of Bcl-2 expression in the central nervous system of aged Fisher-344 rats. J Neurochem. 2001 Feb;76(4):1099–1108. doi: 10.1046/j.1471-4159.2001.00118.x. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Lindeman G. J., Gaubatz S., Livingston D. M., Ginsberg D. The subcellular localization of E2F-4 is cell-cycle dependent. Proc Natl Acad Sci U S A. 1997 May 13;94(10):5095–5100. doi: 10.1073/pnas.94.10.5095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lundberg A. S., Hahn W. C., Gupta P., Weinberg R. A. Genes involved in senescence and immortalization. Curr Opin Cell Biol. 2000 Dec;12(6):705–709. doi: 10.1016/s0955-0674(00)00155-1. [DOI] [PubMed] [Google Scholar]
  22. Marvel J., Perkins G. R., Lopez Rivas A., Collins M. K. Growth factor starvation of bcl-2 overexpressing murine bone marrow cells induced refractoriness to IL-3 stimulation of proliferation. Oncogene. 1994 Apr;9(4):1117–1122. [PubMed] [Google Scholar]
  23. Meikrantz W., Gisselbrecht S., Tam S. W., Schlegel R. Activation of cyclin A-dependent protein kinases during apoptosis. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3754–3758. doi: 10.1073/pnas.91.9.3754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Meikrantz W., Schlegel R. Apoptosis and the cell cycle. J Cell Biochem. 1995 Jun;58(2):160–174. doi: 10.1002/jcb.240580205. [DOI] [PubMed] [Google Scholar]
  25. Mikami T., Yanagisawa N., Baba H., Koike M., Okayasu I. Association of Bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis. Cancer. 1999 Jan 15;85(2):318–325. [PubMed] [Google Scholar]
  26. Mok C. L., Gil-Gómez G., Williams O., Coles M., Taga S., Tolaini M., Norton T., Kioussis D., Brady H. J. Bad can act as a key regulator of T cell apoptosis and T cell development. J Exp Med. 1999 Feb 1;189(3):575–586. doi: 10.1084/jem.189.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Murphy K. L., Kittrell F. S., Gay J. P., Jäger R., Medina D., Rosen J. M. Bcl-2 expression delays mammary tumor development in dimethylbenz(a)anthracene-treated transgenic mice. Oncogene. 1999 Nov 11;18(47):6597–6604. doi: 10.1038/sj.onc.1203099. [DOI] [PubMed] [Google Scholar]
  28. Nicoletti I., Migliorati G., Pagliacci M. C., Grignani F., Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271–279. doi: 10.1016/0022-1759(91)90198-o. [DOI] [PubMed] [Google Scholar]
  29. O'Reilly L. A., Harris A. W., Tarlinton D. M., Corcoran L. M., Strasser A. Expression of a bcl-2 transgene reduces proliferation and slows turnover of developing B lymphocytes in vivo. J Immunol. 1997 Sep 1;159(5):2301–2311. [PubMed] [Google Scholar]
  30. O'Reilly L. A., Huang D. C., Strasser A. The cell death inhibitor Bcl-2 and its homologues influence control of cell cycle entry. EMBO J. 1996 Dec 16;15(24):6979–6990. [PMC free article] [PubMed] [Google Scholar]
  31. Pietenpol J. A., Papadopoulos N., Markowitz S., Willson J. K., Kinzler K. W., Vogelstein B. Paradoxical inhibition of solid tumor cell growth by bcl2. Cancer Res. 1994 Jul 15;54(14):3714–3717. [PubMed] [Google Scholar]
  32. Schindowski K., Leutner S., Müller W. E., Eckert A. Age-related changes of apoptotic cell death in human lymphocytes. Neurobiol Aging. 2000 Sep-Oct;21(5):661–670. doi: 10.1016/s0197-4580(00)00171-8. [DOI] [PubMed] [Google Scholar]
  33. Serrano M., Blasco M. A. Putting the stress on senescence. Curr Opin Cell Biol. 2001 Dec;13(6):748–753. doi: 10.1016/s0955-0674(00)00278-7. [DOI] [PubMed] [Google Scholar]
  34. Shan B., Lee W. H. Deregulated expression of E2F-1 induces S-phase entry and leads to apoptosis. Mol Cell Biol. 1994 Dec;14(12):8166–8173. doi: 10.1128/mcb.14.12.8166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sherr C. J., Roberts J. M. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev. 1999 Jun 15;13(12):1501–1512. doi: 10.1101/gad.13.12.1501. [DOI] [PubMed] [Google Scholar]
  36. Shiyanov P., Hayes S., Chen N., Pestov D. G., Lau L. F., Raychaudhuri P. p27Kip1 induces an accumulation of the repressor complexes of E2F and inhibits expression of the E2F-regulated genes. Mol Biol Cell. 1997 Sep;8(9):1815–1827. doi: 10.1091/mbc.8.9.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stein G. H., Drullinger L. F., Robetorye R. S., Pereira-Smith O. M., Smith J. R. Senescent cells fail to express cdc2, cycA, and cycB in response to mitogen stimulation. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11012–11016. doi: 10.1073/pnas.88.24.11012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tsugu A., Sakai K., Dirks P. B., Jung S., Weksberg R., Fei Y. L., Mondal S., Ivanchuk S., Ackerley C., Hamel P. A. Expression of p57(KIP2) potently blocks the growth of human astrocytomas and induces cell senescence. Am J Pathol. 2000 Sep;157(3):919–932. doi: 10.1016/S0002-9440(10)64605-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vail M. E., Pierce R. H., Fausto N. Bcl-2 delays and alters hepatic carcinogenesis induced by transforming growth factor alpha. Cancer Res. 2001 Jan 15;61(2):594–601. [PubMed] [Google Scholar]
  40. Vairo G., Innes K. M., Adams J. M. Bcl-2 has a cell cycle inhibitory function separable from its enhancement of cell survival. Oncogene. 1996 Oct 3;13(7):1511–1519. [PubMed] [Google Scholar]
  41. Vairo G., Soos T. J., Upton T. M., Zalvide J., DeCaprio J. A., Ewen M. E., Koff A., Adams J. M. Bcl-2 retards cell cycle entry through p27(Kip1), pRB relative p130, and altered E2F regulation. Mol Cell Biol. 2000 Jul;20(13):4745–4753. doi: 10.1128/mcb.20.13.4745-4753.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wang E. Senescent human fibroblasts resist programmed cell death, and failure to suppress bcl2 is involved. Cancer Res. 1995 Jun 1;55(11):2284–2292. [PubMed] [Google Scholar]
  43. Wang Y., Blandino G., Givol D. Induced p21waf expression in H1299 cell line promotes cell senescence and protects against cytotoxic effect of radiation and doxorubicin. Oncogene. 1999 Apr 22;18(16):2643–2649. doi: 10.1038/sj.onc.1202632. [DOI] [PubMed] [Google Scholar]
  44. Wang Y., Blandino G., Oren M., Givol D. Induced p53 expression in lung cancer cell line promotes cell senescence and differentially modifies the cytotoxicity of anti-cancer drugs. Oncogene. 1998 Oct 15;17(15):1923–1930. doi: 10.1038/sj.onc.1202113. [DOI] [PubMed] [Google Scholar]
  45. Zhou B. B., Li H., Yuan J., Kirschner M. W. Caspase-dependent activation of cyclin-dependent kinases during Fas-induced apoptosis in Jurkat cells. Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6785–6790. doi: 10.1073/pnas.95.12.6785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. de La Coste A., Mignon A., Fabre M., Gilbert E., Porteu A., Van Dyke T., Kahn A., Perret C. Paradoxical inhibition of c-myc-induced carcinogenesis by Bcl-2 in transgenic mice. Cancer Res. 1999 Oct 1;59(19):5017–5022. [PubMed] [Google Scholar]
  47. van den Heuvel S., Harlow E. Distinct roles for cyclin-dependent kinases in cell cycle control. Science. 1993 Dec 24;262(5142):2050–2054. doi: 10.1126/science.8266103. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES