Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 2000 Oct;83(8):1039–1046. doi: 10.1054/bjoc.2000.1411

Retinoblastoma protein-initiated cellular growth arrest overcomes the ability of cotransfected wild-type p53 to induce apoptosis

H Shinohara 1,2, J Zhou 3, K Yoshikawa 1,2, S Yazumi 1, K Ko 1, Y Yamaoka 2, T Mizukami 4, T Yoshida 4, S Akinaga 4, T Tamaoki 4, H Motoda 1, W F Benedict 3, R Takahashi 1
PMCID: PMC2363552  PMID: 10993652

Abstract

The retinoblastoma gene, RB, participates in the regulation of the G1/S-phase transition and in p53-mediated apoptosis. We have previously reported that stably transfected RB functions as a growth and tumour suppressor in HTB9 human bladder carcinoma cells, which carry a mutation of the p53 gene at codon 280 and lack RB expression. To elucidate the potential role of RB in the regulation of p53-mediated apoptosis, we transfected a wt p53 expression plasmid under the control of the human cytomegalovirus promoter into parental and RB-transfected HTB9 cells. The p53+/RB– cells were susceptible to apoptosis under various experimental conditions: 1) incubation in serum-free culture for 72 h, 2) short-term (6 h) or long-term (48 h) exposure to etoposide, and 3) culturing in soft agar. In contrast, p53+/RB+ cells were significantly resistant to apoptosis under similar conditions and exhibited efficient growth arrest, as measured by laser scanning cytometry. Tumorigenicity in nude mice of parental HTB9 cells was lost by exogenous expression of wt p53. Likewise, none of mice injected subcutaneously with either p53–/RB+ or p53+/RB+ cells developed tumours, indicating that RB allows suppression of tumorigenesis, regardless of p53 status. These results suggest that the growth-inhibitory function of RB may overcome the ability of wt p53 to induce apoptosis. © 2000 Cancer Research Campaign

Keywords: retinoblastoma gene, p53 gene, transfection, apoptosis, cell cycle

Full Text

The Full Text of this article is available as a PDF (440.6 KB).

Footnotes

Correspondence to: R Takahashi

Selected References

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

  1. Almasan A., Yin Y., Kelly R. E., Lee E. Y., Bradley A., Li W., Bertino J. R., Wahl G. M. Deficiency of retinoblastoma protein leads to inappropriate S-phase entry, activation of E2F-responsive genes, and apoptosis. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5436–5440. doi: 10.1073/pnas.92.12.5436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bartek J., Bartkova J., Lukas J. The retinoblastoma protein pathway in cell cycle control and cancer. Exp Cell Res. 1997 Nov 25;237(1):1–6. doi: 10.1006/excr.1997.3776. [DOI] [PubMed] [Google Scholar]
  3. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. DeGregori J., Leone G., Miron A., Jakoi L., Nevins J. R. Distinct roles for E2F proteins in cell growth control and apoptosis. Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7245–7250. doi: 10.1073/pnas.94.14.7245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Del Bino G., Darzynkiewicz Z. Camptothecin, teniposide, or 4'-(9-acridinylamino)-3-methanesulfon-m-anisidide, but not mitoxantrone or doxorubicin, induces degradation of nuclear DNA in the S phase of HL-60 cells. Cancer Res. 1991 Feb 15;51(4):1165–1169. [PubMed] [Google Scholar]
  7. Haapajärvi T., Kivinen L., Pitkänen K., Laiho M. Cell cycle dependent effects of u.v.-radiation on p53 expression and retinoblastoma protein phosphorylation. Oncogene. 1995 Jul 6;11(1):151–159. [PubMed] [Google Scholar]
  8. Hartwell L. H., Kastan M. B. Cell cycle control and cancer. Science. 1994 Dec 16;266(5192):1821–1828. doi: 10.1126/science.7997877. [DOI] [PubMed] [Google Scholar]
  9. Haupt Y., Rowan S., Oren M. p53-mediated apoptosis in HeLa cells can be overcome by excess pRB. Oncogene. 1995 Apr 20;10(8):1563–1571. [PubMed] [Google Scholar]
  10. Kamentsky L. A., Kamentsky L. D. Microscope-based multiparameter laser scanning cytometer yielding data comparable to flow cytometry data. Cytometry. 1991;12(5):381–387. doi: 10.1002/cyto.990120502. [DOI] [PubMed] [Google Scholar]
  11. Kowalik T. F., DeGregori J., Leone G., Jakoi L., Nevins J. R. E2F1-specific induction of apoptosis and p53 accumulation, which is blocked by Mdm2. Cell Growth Differ. 1998 Feb;9(2):113–118. [PubMed] [Google Scholar]
  12. Kowalik T. F., DeGregori J., Schwarz J. K., Nevins J. R. E2F1 overexpression in quiescent fibroblasts leads to induction of cellular DNA synthesis and apoptosis. J Virol. 1995 Apr;69(4):2491–2500. doi: 10.1128/jvi.69.4.2491-2500.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Macleod K. F., Hu Y., Jacks T. Loss of Rb activates both p53-dependent and independent cell death pathways in the developing mouse nervous system. EMBO J. 1996 Nov 15;15(22):6178–6188. [PMC free article] [PubMed] [Google Scholar]
  14. Morgenbesser S. D., Williams B. O., Jacks T., DePinho R. A. p53-dependent apoptosis produced by Rb-deficiency in the developing mouse lens. Nature. 1994 Sep 1;371(6492):72–74. doi: 10.1038/371072a0. [DOI] [PubMed] [Google Scholar]
  15. Pan H., Yin C., Dyson N. J., Harlow E., Yamasaki L., Van Dyke T. Key roles for E2F1 in signaling p53-dependent apoptosis and in cell division within developing tumors. Mol Cell. 1998 Sep;2(3):283–292. doi: 10.1016/s1097-2765(00)80273-7. [DOI] [PubMed] [Google Scholar]
  16. Pomerantz J., Schreiber-Agus N., Liégeois N. J., Silverman A., Alland L., Chin L., Potes J., Chen K., Orlow I., Lee H. W. The Ink4a tumor suppressor gene product, p19Arf, interacts with MDM2 and neutralizes MDM2's inhibition of p53. Cell. 1998 Mar 20;92(6):713–723. doi: 10.1016/s0092-8674(00)81400-2. [DOI] [PubMed] [Google Scholar]
  17. Qin X. Q., Livingston D. M., Kaelin W. G., Jr, Adams P. D. Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10918–10922. doi: 10.1073/pnas.91.23.10918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Resnicoff M., Burgaud J. L., Rotman H. L., Abraham D., Baserga R. Correlation between apoptosis, tumorigenesis, and levels of insulin-like growth factor I receptors. Cancer Res. 1995 Sep 1;55(17):3739–3741. [PubMed] [Google Scholar]
  19. Shaw P., Bovey R., Tardy S., Sahli R., Sordat B., Costa J. Induction of apoptosis by wild-type p53 in a human colon tumor-derived cell line. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4495–4499. doi: 10.1073/pnas.89.10.4495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Takahashi R., Hashimoto T., Xu H. J., Hu S. X., Matsui T., Miki T., Bigo-Marshall H., Aaronson S. A., Benedict W. F. The retinoblastoma gene functions as a growth and tumor suppressor in human bladder carcinoma cells. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5257–5261. doi: 10.1073/pnas.88.12.5257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tamura T., Aoyama N., Saya H., Haga H., Futami S., Miyamoto M., Koh T., Ariyasu T., Tachi M., Kasuga M. Induction of Fas-mediated apoptosis in p53-transfected human colon carcinoma cells. Oncogene. 1995 Nov 16;11(10):1939–1946. [PubMed] [Google Scholar]
  22. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tsai K. Y., Hu Y., Macleod K. F., Crowley D., Yamasaki L., Jacks T. Mutation of E2f-1 suppresses apoptosis and inappropriate S phase entry and extends survival of Rb-deficient mouse embryos. Mol Cell. 1998 Sep;2(3):293–304. doi: 10.1016/s1097-2765(00)80274-9. [DOI] [PubMed] [Google Scholar]
  24. Waldman T., Lengauer C., Kinzler K. W., Vogelstein B. Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21. Nature. 1996 Jun 20;381(6584):713–716. doi: 10.1038/381713a0. [DOI] [PubMed] [Google Scholar]
  25. White E., Cipriani R., Sabbatini P., Denton A. Adenovirus E1B 19-kilodalton protein overcomes the cytotoxicity of E1A proteins. J Virol. 1991 Jun;65(6):2968–2978. doi: 10.1128/jvi.65.6.2968-2978.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wyllie F. S., Haughton M. F., Bond J. A., Rowson J. M., Jones C. J., Wynford-Thomas D. S phase cell-cycle arrest following DNA damage is independent of the p53/p21(WAF1) signalling pathway. Oncogene. 1996 Mar 7;12(5):1077–1082. [PubMed] [Google Scholar]
  27. Yamasaki L., Bronson R., Williams B. O., Dyson N. J., Harlow E., Jacks T. Loss of E2F-1 reduces tumorigenesis and extends the lifespan of Rb1(+/-)mice. Nat Genet. 1998 Apr;18(4):360–364. doi: 10.1038/ng0498-360. [DOI] [PubMed] [Google Scholar]
  28. Yamasaki L. Growth regulation by the E2F and DP transcription factor families. Results Probl Cell Differ. 1998;22:199–227. doi: 10.1007/978-3-540-69686-5_9. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES