Summary
The G1 cell cycle checkpoint regulates entry into S phase for normal cells. Components of the G1 checkpoint, including retinoblastoma (Rb) protein, cyclin D1 and p16INK4a, are commonly altered in human malignancies, abrogating cell cycle control. Using immunohistochemistry, we examined 79 invasive transitional cell carcinomas of the urinary bladder treated by cystectomy, for loss of Rb or p16INK4a protein and for cyclin D1 overexpression. As p53 is also involved in cell cycle control, its expression was studied as well. Rb protein loss occurred in 23/79 cases (29%); it was inversely correlated with loss of p16INK4a, which occurred in 15/79 cases (19%). One biphenotypic case, with Rb+p16– and Rb-p16+ areas, was identified as well. Cyclin D1 was overexpressed in 21/79 carcinomas (27%), all of which retained Rb protein. Fifty of 79 tumours (63%) showed aberrant accumulation of p53 protein; p53 staining did not correlate with Rb, p16INK4a, or cyclin D1 status. Overall, 70% of bladder carcinomas showed abnormalities in one or more of the intrinsic proteins of the G1 checkpoint (Rb, p16INK4a and cyclin D1). Only 15% of all bladder carcinomas (12/79) showed a normal phenotype for all four proteins. In a multivariate survival analysis, cyclin D1 overexpression was linked to less aggressive disease and relatively favourable outcome. In our series, Rb, p16INK4a and p53 status did not reach statistical significance as prognostic factors. In conclusion, G1 restriction point defects can be identified in the majority of bladder carcinomas. Our findings support the hypothesis that cyclin D1 and p16INK4a can cooperate to dysregulate the cell cycle, but that loss of Rb protein abolishes the G1 checkpoint completely, removing any selective advantage for cells that alter additional cell cycle proteins.
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Aagaard, L., Lukas, J., Bartkova, J., Kjerulff, A-A, Strauss, M. & Bartek, J. (1995). Aberrations of p16INK4 and retinoblastoma tumour-suppressor genes occur in distinct sub-sets of human cancer cell lines. Int J Cancer 61: 115–120.
American Joint Committee on Cancer (1992). Manual for Staging of Cancer, 4th edn.Lippincott: Philadelphia 195–200.
Bartkova, J., Lukas, J., Guldberg, P., Alsner, J., Kirkin, A. F., Zeuthen, J. & Bartek, J. (1996). The p16-cyclin D/Cdk4-pRb pathway as a functional unit frequently altered in melanoma pathogenesis. Cancer Res 56: 5475–5483.
Bringuier, P. P., Tamimi, Y., Schuuring, E. & Schalken, J. (1996). Expression of cyclin D1 and EMS1 in bladder tumours; relationship with chromosome 11q13 amplification. Oncogene 12: 1747–1753.
Cordon-Cardo, C., Wartinger, D., Petrylak, D., Dalbagni, G., Fair, W. R., Fuks, Z. & Reuter, V. E. (1992). Altered expression of the retinoblastoma gene product: prognostic indicator in bladder cancer. J Natl Cancer Inst 84: 1251–1256.
Cordon-Cardo, C., Dalbagni, G., Saez, G. T., Oliva, M. R., Zhang, Z-F, Rosai, J., Reuter, V. E. & Pellicer, A. (1994). p53 mutations in human bladder cancer: genotypic versus phenotypic patterns. Int J Cancer 56: 347–353.
Cote, R. J., Dunn, M. D., Chatterjee, S. J., Stein, J. P., Shi, S-R, Tran, Q-C, Hu, S. X., Xu, H. J., Groshen, S., Taylor, C. R., Skinner, D. G. & Benedict, W. F. (1998). Elevated and absent pRb expression is associated with bladder cancer progression and has cooperative effects with p53. Cancer Res 58: 1090–1094.
El-Deiry, W. S., Tokino, T., Velculescu, V. E., Levy, D. B., Parsons, R., Trent, J. M., Lin, D., Mercer, W. E., Kinzler, K. W. & Vogelstein, B. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.
Esrig, D., Elmajian, D., Groshen, S., Freeman, J. A., Stein, J. P., Chen, S-C, Nichols, P. W., Skinner, D. G., Jones, P. A. & Cote, R. J. (1994). Accumulation of nuclear p53 and tumour progression in bladder cancer. N Engl J Med 331: 1259–1264.
Finlay, C. A., Hinds, P. W., Tan, T-H, Eliyahu, D., Oren, M. & Levine, A. J. (1988). Activating mutations for transformation by p53 produce a gene product that forms an hsc70-p53 complex with an altered half-life. Mol Cell Biol 8: 531–539.
Geradts, J., Kratzke, R. A., Niehans, G. A. & Lincoln, C. E. (1995). Immunohistochemical detection of the cyclin-dependent kinase inhibitor 2/ multiple tumor suppressor gene 1 (CDKN2/MTS1) product p16INK4a in archival human solid tumors: correlation with retinoblastoma protein expression. Cancer Res 55: 6006–6011.
Geradts, J., Kratzke, R. A., Crush-Stanton, S., Wen, S. F. & Lincoln, C. E. (1996). Wild-type and mutant retinoblastoma protein in paraffin sections. Mod Pathol 9: 339–347.
Gillett, C., Smith, P., Gregory, W., Richards, M., Millis, R., Peters, G. & Barnes, D. (1996). Cyclin D1 and prognosis in human breast cancer. Int J Cancer (Pred Oncol) 69: 92–99.
Glick, S. H., Howell, L. P. & White, R. W. D. (1996). Relationship of p53 and bcl-2 to prognosis in muscle-invasive transitional cell carcinoma of the bladder. J Urol 155: 1754–1757.
Gruis, N. A., Weaver-Feldhaus, J., Liu, Q., Frye, C., Eeles, R., Orlow, I., Lacombe, L., Ponce-Castaneda, V., Lianes, P., Latres, E., Skolnick, M., Cordon-Cardo, C. & Kamb, A. (1995). Genetic evidence in melanoma and bladder cancers that p16 and p53 function in separate pathways of tumor suppression. Am J Pathol 146: 1199–1206.
Harbour, J. W., Lai, S-L, Whang-Peng, J., Gazdar, A. F., Minna, J. D. & Kaye, F. J. (1988). Abnormalities in structure and expression of the human retinoblastoma gene in SCLC. Science 241: 353–357.
Horowitz, J. M., Park, S-H, Bogenmann, E., Cheng, J-C, Yandell, D. W., Kaye, F. J., Minna, J. D., Dryja, T. P. & Weinberg, R. A. (1990). Frequent inactivation of the retinoblastoma anti-oncogene is restricted to a subset of human tumour cells. Proc Natl Acad Sci USA 87: 2775–2779.
Jahnson, S., Risberg, B., Karlsson, M. G., Westman, G., Bergström, R. & Pedersen, J. (1995). p53 and Rb immunostaining in locally advanced bladder cancer: relation to prognostic variables and predictive value for the local response to radical radiotherapy. Eur Urol 28: 135–142.
Jiang, W., Zhang, Y-J, Kahn, S. M., Hollstein, M. C., Santella, R. M., Lu, S-H, Harris, C. C., Montesano, R. & Weinstein, I. B. (1993). Altered expression of the cyclin D1 and retinoblastoma genes in human esophageal cancer. Proc Natl Acad Sci USA 90: 9026–9030.
Kotelnikov, V. M., Coon, JSIV, Mundle, S., Kelanic, S., LaFollette, S., Taylor, S. I. V., Hutchinson, J., Panje, W., Caldarelli, D. D. & Preisler, H. D. (1997). Cyclin D1 expression in squamous cell carcinomas of the head and neck and in oral mucosa in relation to proliferation and apoptosis. Clin Cancer Res 3: 95–101.
Kratzke, R. A., Otterson, G. A., Lincoln, C. E., Ewing, S., Oie, H., Geradts, J. & Kaye, F. J. (1995). Immunohistochemical analysis of the p16-INK4a cyclin-dependent kinase inhibitor in malignant mesothelioma. J Natl Cancer Inst 87: 1870–1875.
Kratzke, R. A., Greatens, T. M., Rubins, J. B., Maddaus, M. A., Niewoehner, D. E., Niehans, G. A. & Geradts, J. (1996). Rb and p16INK4a expression in resected non-small-cell lung tumours. Cancer Res 56: 3415–3420.
Lee, C. C. R., Yamamoto, S., Morimura, K., Wanibuchi, H., Nishisaka, N., Ikemoto, S., Nakatani, T., Wada, S., Kishamoto, T. & Fukushima, S. (1997). Significance of cyclin D1 overexpression in transitional cell carcinomas of the urinary bladder and its correlation with histopathologic features. Cancer 79: 780–789.
Li, Y., Nichols, M. A., Shay, J. W. & Xiong, Y. (1994). Transcriptional repression of the D-type cyclin-dependent kinase kinase inhibitor p16 by the retinoblastoma susceptibility gene product pRb. Cancer Res 54: 6078–6082.
Lipponen, P. K. (1993). Over-expression of p53 nuclear oncoprotein in transitional cell bladder cancer and its prognostic value. Int J Cancer 53: 365–370.
Lipponen, P. K. & Liukkonen, T. J. O. (1995). Reduced expression of retinoblastoma (Rb) gene protein is related to cell proliferation and prognosis in transitional-cell bladder cancer. J Cancer Res Clin Oncol 121: 44–50.
Logothetis, C. J., Xu, H-J, Ro, J. Y., Hu, S-X, Sahin, A., Ordonez, N. & Benedict, W. F. (1992). Altered expression of retinoblastoma protein and known prognostic variables in locally advanced bladder cancer. J Natl Cancer Inst 84: 1256–1261.
Lukas, J., Aagaard, L., Strauss, M. & Bartek, J. (1995a). Oncogenic aberrations of p16INK4/CDKN2 and cyclin D1 cooperate to deregulate G1 control. Cancer Res 55: 4818–4823.
Lukas, J., Bartkova, J., Rohde, M., Strauss, M. & Bartek, J. (1995b). Cyclin D1 is dispensable for G1 control in retinoblastoma gene-deficient cells independently of cdk4 activity. Mol Cell Biol 15: 2600–2611.
Lukas, J., Parry, D., Aagard, L., Mann, D. J., Bartkova, J., Strauss, M., Peters, G. & Bartek, J. (1995c). Retinoblastoma-protein-dependent cell-cycle inhibition by the tumour suppressor p16. Nature 375: 503–506.
Medema, R. H., Herrera, R. E., Lam, F. & Weinberg, R. A. (1995). Growth suppression by p16-ink4 requires functional retinoblastoma protein. Proc Natl Acad Sci USA 92: 6289–6293.
Nguyen, P. L., Swanson, P. E., Jaszcz, W., Aeppli, D. M., Zhang, G., Singleton, T. P., Ward, S., Dykoski, D., Harvey, J. & Niehans, G. A. (1994). Expression of epidermal growth factor receptor in invasive transitional cell carcinoma of the urinary bladder: A multivariate survival analysis. Am J Clin Pathol 101: 166–176.
Okamoto, A., Demetrick, D. J., Spillare, E. A., Hagiwara, K., Hussain, S. P., Bennett, W. P., Forrester, K., Gerwin, B., Greenblatt, M. S., Serrano, M., Shiseki, M., Yokota, J., Beach, D. H. & Harris, C. C. (1994). p16-INK4 mutations and altered expression in human tumors and cell lines. Cold Spring Harbor Symp Quant Biol 59: 49–57.
Orlow, I., Lacombe, L., Hannon, G. J., Serrano, M., Pellicer, I., Dalbagni, G., Reuter, V. E., Zhang, Z-F, Beach, D. & Cordon-Cardo, C. (1995). Deletion of the p16 and p15 genes in human bladder tumors. J Natl Cancer Inst 87: 1524–1529.
Otterson, G. A., Kratzke, R. A., Coxon, A., Kim, Y. W. & Kaye, F. J. (1994). Absence of p16INK4 protein is restricted to the subset of lung cancer lines that retains wildtype RB. Oncogene 9: 3375–3378.
Packenham, J. P., Taylor, J. A., Anna, C. H., White, C. M. & Devereux, T. (1995). Homozygous deletions but no sequence mutations in coding regions of p15 or p16 in human primary bladder tumours. Mol Carcinogen 14: 147–151.
Parry, D., Bates, S., Mann, D. J. & Peters, G. (1995). Lack of cyclin D-cdk complexes in Rb-negative cells correlates with high levels of p16INK4/MTS1 tumour suppressor gene product. EMBO J 14: 503–511.
Resnick, J. M., Cherwitz, D., Knapp, D., Uhlman, D. & Niehans, G. A. (1995). A microwave method that enhances detection of aberrant p53 expression in formalin-fixed, paraffin-embedded tissues. Arch Pathol Lab Med 119: 360–366.
Rosai, J. (1996). Ackerman’s Surgical Pathology, 8th edn, pp. 1197–1201. Mosby: St Louis
Sakaguchi, M., Fujii, Y., Hirabayashi, H., Yoon, H-E, Komoto, Y., Oue, T., Kusafuka, T., Okada, A. & Matsuda, H. (1996). Inversely correlated expression of p16 and Rb protein in non-small-cell lung cancers: an immunohistochemical study. Int J Cancer 65: 442–445.
Sarkis, A. S., Dalbagni, G., Cordon-Cardo, C., Zhang, Z-F, Sheinfeld, J., Fair, W. R., Herr, H. W. & Reuter, V. E. (1993). Nuclear overexpression of p53 protein in transitional cell bladder carcinoma: a marker for disease progression. J Natl Cancer Inst 85: 53–59.
Schauer, I. E., Siriwardana, S., Langan, T. A. & Sclafani, R. A. (1994). Cyclin D1 overexpression vs. retinoblastoma inactivation: implications for growth control evasion in non-small cell and small cell lung cancer. Proc Natl Acad Sci USA 91: 7827–7831.
Schutte, M., Hruban, R. H., Geradts, J., Maynard, R., Hilgers, W., Rabindran, S. K., Moskaluk, C. A., Hahn, S. A., Schwarte-Waldhoff, I., Schmiegel, W., Baylin, S. B., Kern, S. E. & Herman, J. G. (1997). Abrogation of the Rb/p16 tumor-suppressive pathway in virtually all pancreatic carcinomas. Cancer Res 57: 3126–3130.
Serrano, M., Hannon, G. J. & Beach, D. (1993). A new regulatory motif in cell-cycle control causing specific inhibition of cyclinD/CDK4. Nature 366: 704–707.
Shapiro, G. I., Edwards, C. D., Kobzik, L., Godleski, J., Richards, W., Sugarbaker, D. J. & Rollins, B. J. (1995). Reciproca Rb inactivation and p16INK4 expression in primary lung cancers and cell lines. Cancer Res 55: 505–509.
Shin, K. Y., Kong, G., Kim, W. S., Lee, T. Y., Woo, Y. N. & Lee, J. D. (1997). Overexpression of cyclin D1 correlates with early recurrence in superficial bladder cancers. Br J Cancer 75: 1788–1792.
Strauss, M., Lukas, J. & Bartek, J. (1995). Unrestricted cell cycling and cancer. Nature Med 1: 1245–1246.
Tam, S. W., Theodoras, A. M., Shay, J. W., Draetta, G. F. & Pagano, M. (1994). Differential expression and regulation of cyclin D1 protein in normal and tumour human cells: association with Cdk4 is required for cyclin D1 function in G1 progression. Oncogene 9: 2663–2674.
Têtu, B., Fradet, Y., Allard, P., Veilleux, C., Roberge, N. & Bernard, P. (1996). Prevalence and clinical significance of HER-2/neu, p53 and Rb expression in primary superficial bladder cancer. J Urol 155: 1784–1788.
Tsao, H., Benoit, E., Sober, A. J., Thiele, C. & Haluska, F. G. (1998). Novel mutations in the p16/CDKN2A binding region of the cyclin-dependent kinase-4 gene. Cancer Res 58: 109–113.
Ueki, K., Ono, Y., Henson, J. W., Efird, J. T., von Deimling, A. & Louis, D. N. (1996). CDKN2/p16 or RB alterations occur in the majority of glioblastomas and are inversely correlated. Cancer Res 56: 150–153.
Vet, J. A. M., Bringuier, P. P., Schaafsma, H. E., Witjes, J. A., Debruyne, F. M. J. & Schalken, J. A. (1995). Comparison of p53 protein overexpression with p53 mutation in bladder cancer: clinical and biologic aspects. Lab Invest 73: 837–843.
Williamson, M. P., Elder, P. A., Shaw, M. E., Devlin, J. & Knowles, M. A. (1995). p16 (CDKN2) is a major deletion target at 9p21 in bladder cancer. Hum Mol Genet 4: 1569–1577.
Zuo, L., Weger, J., Yang, Q., Goldstein, A. M., Tucker, M. A., Walker, G. J., Hayward, N. & Dracopoli, N. C. (1996). Germline mutations in the p16INK4a binding domain of CDK4 in familial melanoma. Nat Genet 12: 97–99.
Author information
Authors and Affiliations
Additional information
Presented in part at the 87th Annual Meeting of the US and Canadian Academy of Pathology, 28 February to 6 March 1998, in Boston, MA, USA
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Niehans, G., Kratzke, R., Froberg, M. et al. G1 checkpoint protein and p53 abnormalities occur in most invasive transitional cell carcinomas of the urinary bladder. Br J Cancer 80, 1175–1184 (1999). https://doi.org/10.1038/sj.bjc.6990483
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6990483
Keywords
This article is cited by
-
The impact of cyclin D1 overexpression on the prognosis of bladder cancer: a meta-analysis
World Journal of Surgical Oncology (2014)
-
Clinical significance of altered nm23-H1, EGFR, RB and p53 expression in bilharzial bladder cancer
BMC Cancer (2009)
-
Prognostic value of cell cycle regulatory proteins in muscle-infiltrating bladder cancer
Journal of Cancer Research and Clinical Oncology (2006)
-
Human papilloma virus and p53 expression in bladder cancer in Egypt: Relationship to schistosomiasis and clinicopathologic factors
Pathology & Oncology Research (2006)
-
Expression of cyclin D1 in normal, hyperplastic and neoplastic endometrium and its correlation with Ki-67 and clinicopathological variables
Archives of Gynecology and Obstetrics (2005)