Cellular Senescence and the Cell Cycle

  • J. Carl Barrett
  • Cynthia A. Afshari
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)


Normal cells in culture can be grown for only a limited number of cell divisions after which they exhibit morphological changes and cease proliferation, a process termed cellular senescence or cellular aging (1). Hayflick and Moorhead (2) reported this finding with human fibroblasts over 30 years ago, and it has been subsequently confirmed by many investigators using cells from different tissues and species. The failure of cells to grow beyond this limit is an inherent property of the cells that cannot be explained simply by inadequate media components or growth conditions (1, 2). The key determinant in the life span of cells in culture is the number of cell doublings, not the length of time in culture (1). Normal cells transplanted serially in vivo also exhibit a finite life span, suggesting that cellular senescence is not a cell culture artifact (3). Several lines of evidence suggest that the aging of cells in culture may be related to the aging of the organism (1, 4). These lines of evidence, although not conclusive, provide provocative support for the hypothesis that aging of cells is related to the aging process of the organism.


Protein Phosphatase Cellular Senescence Okadaic Acid Senescent Cell Senescence Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Hayflick, L. The cell biology of human aging. N. Engl. J. Med. 295: 1302–1308 (1976).PubMedCrossRefGoogle Scholar
  2. 2.
    Hayflick, L. and Moorhead, P.S. The serial cultivation of human diploid cell strains. Exp. Cell Res. 5: 585–621 (1961).CrossRefGoogle Scholar
  3. 3.
    Daniel, C.W., DeOme, K.B., Young, J.T., Blair, P.B., and Faulkin, L.J., Jr. The in vivo span of normal and preneoplastic mouse mammary glands: a serial transplantation study. Proc. Nati. Acad. Sci. USA 61: 53–60 (1968).CrossRefGoogle Scholar
  4. 4.
    Barrett, J.C. Cell Senescence and Apoptosis. Molecular Genetics of Nervous System Tumors. 61–72 (1993).Google Scholar
  5. 5.
    Barrett, J.C. and Fletcher, W.F. Cellular and molecular mechanisms of multistep carcinogenesis in cell culture models, in: “Mechanisms of Environmental Carcinogenesis: Multistep Models of Carcinogenesis,” Volume II, J. C. Barrett, ed., CRC Press, Boca Raton, 1987.Google Scholar
  6. 6.
    Sager, R. Genetic suppression of tumor formation: a new frontier in cancer research. Cancer Res. 46: 1573–1580 (1986).PubMedGoogle Scholar
  7. 7.
    Maciera-Coelho, A. Biology of normal proliferating cells in vitro. Relevance for in vivo aging, in: “Interdisciplinary Topics in Gerontology,” Volume 23, H.P. von Hang, ed. Karger, Basel (1988).Google Scholar
  8. 8.
    Pereira-Smith, O.M. and Smith, J.R. Genetic analysis of indefinite division in human cells: Identification of four complementation groups. Proc. Natl. Acad. Sci. USA 85: 6042–6046 (1988).PubMedCrossRefGoogle Scholar
  9. 9.
    Sugawara, O.M., Oshimura, M., Koi, M., Annab, L., and Barrett, J.C. Induction of cellular senescence in immortalized cells by human chromosome 1. Science 247: 707–710 (1990).PubMedCrossRefGoogle Scholar
  10. 10.
    Koi, M. and Barrett, J.C. Loss of tumor-suppressive function during chemically induced neoplastic progression of Syrian hamster embryo cells. Proc. Natl. Acad. Sci. USA 83: 5992–5996 (1986).PubMedCrossRefGoogle Scholar
  11. 11.
    Bunn, C.L., and Tarrant, G.M. Limited lifespan in somatic cell hybrids and cybrids Evp. Cell Res. 127: 385–396 (1980).CrossRefGoogle Scholar
  12. 12.
    Klein, C.B., Conway, K., Wang, X.W., Bhamra, R.K., Lin, X., Cohen, M.D., Annab, L., Barrett, J.C., and Costa, M. Senescence of nickel-transformed cells by a mammalian X chromosome: possible epigenetic control. Science 251: 796–799 (1991).PubMedCrossRefGoogle Scholar
  13. 13.
    Ning, Y., Weber, J.L., Ki11, A.M., Ledbetter, D.H., Smith, J.R., and Pereira-Smith, O.M. Genetic analysis of indefinite division in human cells: Evidence for a senescence-related gene (s) on human chromosome 4. Proc. Natl. Acad. Sci. USA 88: 5635–5639 (1991).PubMedCrossRefGoogle Scholar
  14. 14.
    Hara, E., Tsurui, H., Shinozaki, A., Nakada, S., and Oda, K. Cooperative effect of antisense-Rb and antisense-p53 oligomers on the extension of life span in human diploid fibroblasts, TIG-1. Biochem. Biophys. Res. Commun. 179: 528–534 (1991.PubMedCrossRefGoogle Scholar
  15. 15.
    Hinds, P.W., Mittnacht, S., Dulic, V., Arnold, A. Reed, S.I., and Weinberg, R.A. Regulation of retinoblastoma protein functions by ectopic expression of human cyclins. Cell 70: 993–1006 (1992).PubMedCrossRefGoogle Scholar
  16. 16.
    Levine, A.J., Momand, J., and Finlay, C.A. The p53 tumor suppressor gene. Nature 351: 453–456 (1991).PubMedCrossRefGoogle Scholar
  17. 17.
    Goto, M., Rubenstein, M., Weber, J., Woods, K., and Drayna, D. Genetic linkage of Werner’s syndrome to five markers on chromosome 8. Nature 355: 735–738 (1992).PubMedCrossRefGoogle Scholar
  18. 18.
    Sasaki, M., Honda, T., Yamada, H., Wake, N., Barrett, J.C., and Oshimura, M. Evidence for multiple pathways to cellular senescence. Submitted.Google Scholar
  19. 19.
    Wright, W.E., and Shay, J.W. The two-stage mechanism controlling cellular senescence and immortalization. Exp. Gerontol. 27: 383–389 (1992).PubMedCrossRefGoogle Scholar
  20. 20.
    Hubbard-Smith, K., Patsalis, P., Pardinas, J.R., Jha, K.K., Henderson, A.S., and Ozer, H.L. Altered chromosome 6 in immortal human fibroblasts. Mol. Cell. Biol. 12: 2273 (1992).PubMedGoogle Scholar
  21. 21.
    Duncan, E.L., Whitaker, N.J., Moy, E.L., and Reddel, R.R. Assignment of SV40-immortalized cells to more than one complementation group for immortalization. Exp. Cell Res. 205: 337–344 (1993).PubMedCrossRefGoogle Scholar
  22. 21.
    Duncan, E.L., Whitaker, N.J., Moy, E.L., and Reddel, R.R. Assignment of SV40-immortalized cells to more than one complementation group for immortalization. Exp. Cell Res. 205: 337–344 (1993).PubMedCrossRefGoogle Scholar
  23. 23.
    Paraskeva, C., Finarty, S., and Powell, S. Immortalization of a human colorectal adenoma cell line by continuous in vitro passage: possible involvement of chromosome 1 in tumour progression. Int. J. Cancer 41: 908–912 (1988).PubMedCrossRefGoogle Scholar
  24. 24.
    Paraskeva, C., Finarty, S., Mountford, R.A., and Powell, S.C. Specific cytogenetic abnormalities in two new human colorectal adenoma-derived epithelial cell Lines. Cancer Res. 49: 1282–1286 (1989).PubMedGoogle Scholar
  25. 25.
    Paraskeva, C., Harvey, A., Finarty, S., and Powell, S. Possible involvement of chromosome 1 in in vitro immortalization: evidence from progression of a human adenoma-derived cell line in vitro. Int. J. Cancer 43: 743–746 (1989).PubMedCrossRefGoogle Scholar
  26. 26.
    Seshadri, T. and Campisi, J. 1990. Repression of c-fos transcription and an altered genetic program in senescent human fibroblasts. Science 247: 205–209 (1990).Google Scholar
  27. 27.
    Riabowol, K., Schiff, J., and Gilman, M.Z. Transcription factor AP-1 activity is required for initiation of DNA synthesis and is lost during cellular aging. Proc. Natl. Acad. Sci. USA 89: 157–161 (1992).PubMedCrossRefGoogle Scholar
  28. 28.
    Afshari, C.A., Bivins, H.B., and Barrett, J.C. Utilization of a fos-lacZ plasmid to investigate the activation of c fos during cellular senescence and okadaic acid-induced apoptosis. Submitted.Google Scholar
  29. 29.
    Stein, G.H., Beeson, M., and Gordon, L. Failure to phosphorylate the retinoblastoma gene product in senescent human fibroblasts. Science 249: 666–669 (1990).PubMedCrossRefGoogle Scholar
  30. 30.
    Futreal, P.A., and Barrett, J.C. Failure of senescent cells to phosphorylate the RB protein. Oncogen 6: 1109–1113 (1991).Google Scholar
  31. 31.
    Richter, K.H., Afshari, C.A., Annab, L.A., Burkhart, B.A., Owen, R.D., Boyd, J., and Barrett, J.C. Down-regulation of cdc2 in senescent human and hamster cells. Cancer Res. 51: 6010–6013 (1991).PubMedGoogle Scholar
  32. 32.
    Stein, G.H., Drullinger, L.F., Robetorye, R.S., Pereira-Smith, O.M., and Smith, J.R. Senescent cells fail to express cdc2, cycA, and cycB in response to mitogen stimulation. Proc. Natl. Acad. Sct. USA 88: 11012–11016 (1991).CrossRefGoogle Scholar
  33. 33.
    Won, K-A., Xiong, Y., Beach, D., and Gilman, M.Z. Growth regulated expression of D-type cyclin genes in human diploid fibroblasts. Proc. Natl. Acad. Sci. USA 89: 9910–9914 (1992).PubMedCrossRefGoogle Scholar
  34. 34.
    Afshari, C.A., Vojta, P.J. Bivins, H.B., Annab, L.A., Willard, T.B., Futreal, A.F., and Barrett, J.C. Investigation of the role of G, /S cell cycle mediators in cellular senescence. Submitted.Google Scholar
  35. 35.
    Hunter, T. A thousand and one protein kinases. Cell 50: 823–829 (1987).PubMedCrossRefGoogle Scholar
  36. 36.
    Gordon, J.A. Use of vanadate as protein-phosphotyrosine phosphatase inhibitor. Meth. Enrymol. 201: 477–482 (1991).CrossRefGoogle Scholar
  37. 37.
    Cohen, P., Holmes, C.F.B., and Tsukitani, Y. Okadaic acid: a new probe for the study of cellular regulation. Trends Biochem. Sci. 15: 98–102 (1990).PubMedCrossRefGoogle Scholar
  38. 38.
    Honkanen, R.E., Zwiller, J., Daily, S.L., Khatra, B.S., Dukelow, M., and Boynton, A.L. Identification, purification, and characterization of a novel serine/threonine protein phosphatase from bovine brain. J. Biol. Chem. 266: 6614–6619 (1991).PubMedGoogle Scholar
  39. 39.
    Afshari, C.A. and Barrett, J.C. Disruption of G0G1 arrest in quiescent and senescent cells treated with phosphatase inhibitors. Submitted.Google Scholar
  40. 40.
    Afshari, C.A., Kodams, S., Bivins, H.M., Willard, T.B., Fujiki, H., and Barrett, J.C. Induction of neoplastic progression to Syrian hamster embryo cells treated with phosphatase inhibitors. Cancer Res. 53: 1777–1782 (1992).Google Scholar
  41. 41.
    Schdnthal, A., and Feramisco, J.R. Inhibition of histone H1 kinase expression, retinoblastoma protein phosphorylation, and cell proliferation by the phosphatase inhibitor okadaic acid. Oncogen 8: 433–441 (1993).Google Scholar
  42. 42.
    Hardie, D.G., Haystead, T.A.J., and Sim, A.T.R. Use of okadaic acid to inhibit protein phosphatases in intact cells. Meth. Enrymol. 201: 469–476 (1991).CrossRefGoogle Scholar
  43. 43.
    Chen, J., Martin, B.L., and Brautigan, D.L. Regulation of protein serine-threonine phosphatase type 2A by tyrosine phosphorylation. Science 257: 1261–1264 (1992).PubMedCrossRefGoogle Scholar
  44. 44.
    Pallas, D.C., Shahrik, L.K., Martin, B.L., Jaspers, S., Miller, T.B., Brautigan, D.L., and Roberts, T. Polyoma small and middle T antigens and SV40 small t antigen form stable complexes with protein phosphatase 2A. Cell 60: 167–176 (1990).PubMedCrossRefGoogle Scholar
  45. 45.
    Walter, G., Ruediger, R., Slaughter, C., and Mumby, M. 1990. Association of protein phosphatase 2A with polyoms virus medium tumor antigen. Proc. Natla Acad. Sci. USA 87: 2521–2525 (1990).Google Scholar
  46. 46.
    Yang, Lickteig, R.L., Estes, R., Rundell, K., Walter, G., and Mumby, M. Control of protein phosphatase 2A by Simian virus 40 small-t antigen. Mol. Cell. Biol. 11: 1988–1995 (1991).PubMedGoogle Scholar
  47. 47.
    Scheidtmann, K.H., Virshup, D.M., and Kelly, T.J. Protein phosphatase 2A dephosphorylates simian virus 40 large T antigen specifically at residues involved in regulation of DNA-binding activity. J Virol. 65: 2098–2101 (1991).PubMedGoogle Scholar
  48. 48.
    Ide, T., Tsuji, Y., Ishibashi, S., and Mitsui, Y. Reinitiation of host DNA synthesis in senescent human diploid cells by infection with Simian virus 40. Exp. Cell Res. 143: 343–349 (1983).PubMedCrossRefGoogle Scholar
  49. 49.
    Alberts, A.S., Thorbum, A.M., Shenolikar, S., Mumby, M.C., and Feramisco, J.R. Regulation of cell cycle progression and nuclear affinity of the retinoblastoma protein by protein phosphatases. Proc. Natl. Acad. Scl. USA 90: 388–392 (1993).PubMedCrossRefGoogle Scholar
  50. 50.
    Rittling, S.R., Brooks, K.M., Cristofalo, V.J., and Baserga, R. Expression of cell cycle-dependent genes in young and senescent WI-38 fibroblasts. Proc. Natl. Acad. Sci. USA 83: 3316–3320 (1986).PubMedCrossRefGoogle Scholar
  51. 51.
    Yamada, H., Wake, N., Fujimoto, S., Barrett, J.C., and Oshimura, M. Multiple chromosomes carrying tumor suppressor activity for a uterine endometrial carcinoma cell line identified by microcell-mediated chromosome transfer. Oncogene 5: 1141–1447 (1990).PubMedGoogle Scholar
  52. 52.
    Annab, L.A., Barrett, J.C., Hensler, P., and Smith, O., unpublished.Google Scholar
  53. 53.
    Kodams, S., Oshimura, M., and Barrett, J.C. Introduction of a normal human chromosome 8 into SV40-transformed Werner syndrome cells by microcell fusion. Submitted.Google Scholar
  54. 54.
    Ozer, H. Personal communication.Google Scholar
  55. 55.
    Diaz, M. Personal communication.Google Scholar
  56. 56.
    Oshimura, M. Personal communication.Google Scholar
  57. 57.
    Koi, M., Johnson, L.A., Kalikin, L.M., Little, P.F.R., Nakamura, Y., and Feinberg, A.P. Tumor cell growth arrest caused by subchromosomal transferable DNA fragments from chromosome 11. Science 260: 361–364.Google Scholar
  58. 58.
    Lucibello, F.C., Sewing, A., Brüsselbach, S., Bürger, C., and Müller, R. Deregulation of cyclins D1 and E and suppression of cdk2 and cdk4 in senescent human fibroblasts. J Cell Sci. 105: 123–133 (1993).Google Scholar
  59. 59.
    DeVita, V.T. Jr., Young, R.C., and Canellos, G.P. Combination versus single agent chemotherapy: A review of the basis for selection of drug treatment of cancer. Cancer 35: 98–110, 1975.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • J. Carl Barrett
    • 1
  • Cynthia A. Afshari
    • 2
  1. 1.Laboratory of Molecular Carcinogenesis Environmental Carcinogenesis ProgramNational Institute of Environmental Health Sciences National Institutes of HealthResearch Triangle ParkUSA
  2. 2.Center for the Study of Aging and Human DevelopmentDuke University Medical SchoolDurhamUSA

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