Experimental Cancer

Growth arrest and suppression of tumorigenicity of bladder carcinoma cell lines induced by the P16/CDKN2 (p16^INK4A, MTS₁) gene and other loci on human chromosome 9

Qiang Wu

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Laura Possati,

Laura Possati

Institute of Biomedical Sciences, School of Medicine, University of Ancona, Ancona, Italy

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Marco Montesi,

Marco Montesi

Institute of Biomedical Sciences, School of Medicine, University of Ancona, Ancona, Italy

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Francesca Gualandi,

Francesca Gualandi

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Paola Rimessi,

Paola Rimessi

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Cristina Morelli,

Cristina Morelli

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Cecilia Trabanelli,

Cecilia Trabanelli

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Giuseppe Barbanti Brodano,

Corresponding Author

Giuseppe Barbanti Brodano

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

Institute of Microbiology, University of Ferrara, Via Luigi Borsari 46. I-44100 Ferrara, ItalySearch for more papers by this author

Qiang Wu,

Qiang Wu

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Laura Possati,

Laura Possati

Institute of Biomedical Sciences, School of Medicine, University of Ancona, Ancona, Italy

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Marco Montesi,

Marco Montesi

Institute of Biomedical Sciences, School of Medicine, University of Ancona, Ancona, Italy

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Francesca Gualandi,

Francesca Gualandi

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Paola Rimessi,

Paola Rimessi

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Cristina Morelli,

Cristina Morelli

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Cecilia Trabanelli,

Cecilia Trabanelli

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

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Giuseppe Barbanti Brodano,

Corresponding Author

Giuseppe Barbanti Brodano

Institute of Microbiology and Interdepartment Center for Biotechnology, School of Medicine, University of Ferrara, Ferrara

Institute of Microbiology, University of Ferrara, Via Luigi Borsari 46. I-44100 Ferrara, ItalySearch for more papers by this author

First published: 15 March 1996

https://doi.org/10.1002/(SICI)1097-0215(19960315)65:6<840::AID-IJC22>3.0.CO;2-6

Citations: 36

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Abstract

Wild-type P16/CDKN2 (p16^INK4A, MTSI) cDNA, directed by the cytomegalovirus (CMV) immediate early promoter, was transfected into RT4 and RT112 bladder-carcinoma cell lines bearing a mutated endogenous P16/CDKN2 gene and lacking endogenous P16/CDKN2 respectively. In both cases, only transfected clones with rearranged exogenous P16/CDKN2 cDNA could be grown and propagated in cell culture. This result is reminiscent of transfection of wild-type p53 into cells with a deleted or mutated endogenous gene and suggests that P16/CDKN2, over-expressed under control of the strong CMV promoter, induces growth arrest in RT4 and RT112 cells. Transfer of human chromosome 9 to RT4 cells produced RT4/H9 hybrid clones retaining the P16/CDKN2 gene, since in RT4/H9 cell clones P16/CDKN2-gene expression is modulated by the physiological control of chromosomal regulatory sequences. All the RT4/H9 clones lost the entire chromosome 9, except clone 4 and clone 5, which maintained a deleted and an intact chromosome 9 respectively. Loss of several loci in 9p21, including P16/CDKN2, in tumors induced in nude mice by clone 4 and clone 5 suggests that P16/CDKN2 or other genes in 9p21 suppress tumorigenicity in bladder-carcinoma cells. Tumors induced by clone 4 and clone 5 show loss of markers in 9q. The regions 9q22.3, 9q32–33 and 9q34.2, which were maintained in the 2 clones and lost in their derived tumors, may contain tumor-suppressor genes relevant in bladder carcinoma. The results of this study suggest that the P16/CDKN2 gene controls growth of bladder-carcinoma cells when it is over-expressed, and may be involved in the development of bladder carcinoma, but other genes in 9p21 and 9q may participate in bladder-cancer progression. © 1996 Wiley-Liss, Inc.

References

Bassam, B. J., Caetano-Anollés, G. and Gresshoff, P. M., Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal. Biochem., 196, 80–83 (1991).
10.1016/0003-2697(91)90120-I
CAS PubMed Web of Science® Google Scholar
Cairns, P., Mao, L., Merlo, A., Lee, D. J., Schwab, D., Eby, Y., Tokino, K., Van der Riet, P., Blaugrund, J. E. and Sidransky, D., Rates of p16 (p16/CDKN2) mutations in primary tumors with 9p loss. Science, 265, 415–416 (1994).
10.1126/science.8023167
CAS PubMed Web of Science® Google Scholar
Cairns, P. and 20 others, Frequency of homozygous deletion at P16/CDKN2 in primary human tumours. Nature Genet., 11, 210–212 (1995).
10.1038/ng1095-210
CAS PubMed Web of Science® Google Scholar
Cairns, P., Shaw, M. E. and Knowles, M. A., Preliminary mapping of the deleted region of chromosome 9 in bladder cancer. Cancer Res., 53, 1230–1232 (1993).
CAS PubMed Web of Science® Google Scholar
Caldas, C., Hahn, S. A., Da Costa, L. T., Redson, M. S., Schutte, M., Seymour, A. B., Weinstein, C. L., Hruban, R. H., Yeo, C. J. and Kern, S. E., Frequent somatic mutations and homozygous deletions of the p16 (p16/CDKN2) gene in pancreatic adenocarcinoma. Nature Genet., 8, 27–32 (1994).
10.1038/ng0994-27
CAS PubMed Web of Science® Google Scholar
Chaganti, S. R., Gaidano, G., Louie, D. C., Dalla Favera, R. and Chaganti, R. S. K., Diffuse large-Cell lymphomas exhibit frequent deletions in 9p21-22 and 9q31-34 regions. Genes Chrom. Cancer, 12, 32–36 (1995).
10.1002/gcc.2870120106
CAS PubMed Web of Science® Google Scholar
Cordon-Cardo, C., Dalbagni, G., Sarkis, A. S. and Reuter, V. E., Genetic alterations associated with bladder cancer. In: V. T. DeVita, S. Hellman and S. A. Rosenberg (eds.), Important advances in oncology, pp. 71–83, J. B. Lippincott, Philadelphia (1994).
Google Scholar
Devlin, J., Keen, A. J. and Knowles, M. A., Homozygous deletion mapping at 9p21 in bladder carcinoma defines a critical region within 2cM of IFNA. Oncogene, 9, 2757–2760 (1994).
CAS PubMed Web of Science® Google Scholar
Green, A. J., Johnson, P. H. and Yates, J. R. W., The tuberous sclerosis gene on chromosome 9q34 acts as a growth suppressor. Hum. mol. Genet., 3, 1833–1834 (1994).
10.1093/hmg/3.10.1833
CAS PubMed Web of Science® Google Scholar
Habuchi, T., Devlin, J., Elder, P. A. and Knowles, M. A., Detailed deletion mapping of chromosome 9q in bladder cancer: evidence for two tumour suppressor loci. Oncogene, 11, 1671–1674 (1995).
CAS PubMed Web of Science® Google Scholar
Hannon, G. J. and Beach, D., p15^ink4b is a potential effector of TGF-β-Induced cell cycle arrest. Nature (Lond.), 371, 257–261 (1994).
10.1038/371257a0
CAS PubMed Web of Science® Google Scholar
Kamb, A., Gruis, N. A., Weaver-Feldhaus, J., Liu, Q., Harshman, K., Tavtigian, S. V., Stockert, E., Day, R. S., Johnson, B. E. and Skolnick, M. H., A cell cycle regulator potentially involved in genesis of many tumors types. Science, 264, 436–440 (1994).
10.1126/science.8153634
CAS PubMed Web of Science® Google Scholar
Linnenbach, A. J., Presseler, L. B., Seng, B. A., Kimmel, B. S., Tomaszewski, J. E. and Malkowicz, S. B., Characterization of chromosome 9 deletions in transitional cell carcinoma by microsatellite assay. Hum. mol. Genet., 2, 1407–1411 (1993).
10.1093/hmg/2.9.1407
CAS PubMed Web of Science® Google Scholar
Marshall, C. J., Franks, L. M. and Carbonell, A. W., Markers of neoplastic transformation in epithelial cell lines derived from human carcinomas. J. nat. Cancer Inst., 58, 1743–1751 (1977).
10.1093/jnci/58.6.1743
CAS PubMed Web of Science® Google Scholar
Miyao, N., Tsai, Y. C., Lerner, S. P., Olumi, A. F., Spruck III, C. H., Gonzales-Zulueta, M., Nichols, P. W., Skinner, D. G. and Jones, P. A., Role of chromosome 9 in human bladder cancer. Cancer Res., 53, 4066–4070 (1993).
CAS PubMed Web of Science® Google Scholar
Mori, T., Miura, K., Aoki, T., Nishihira, T., Mori, S. and Nakamura, Y., Frequent somatic mutation of the P16/CDKN2/CDK41 (multiple tumor-Suppressor/Cyclin-Dependent kinase 4 inhibitor) gene in esophageal squamous cell carcinoma. Cancer Res., 54, 3396–3397 (1994).
CAS PubMed Web of Science® Google Scholar
Negrini, M., Sabbioni, S., Haldar, S., Possati, L., Castagnoli, A., Corallini, A., Barbanti-Brodano, G. and Croce, C. M., Tumor and growth suppression of breast cancer cells by chromosome 17 associated functions. Cancer Res., 54, 1818–1824 (1994).
CAS PubMed Web of Science® Google Scholar
Nobori, T., Miura, K., Wu, D. J., Lois, A., Takabayashi, K. and Carson, D. A., Deletion of the cyclin-Dependent kinase-4 inhibitor gene in multiple human cancers. Nature (Lond.), 368, 753–756 (1994).
10.1038/368753a0
PubMed Web of Science® Google Scholar
Okamoto, A., Demetrick, D. J., Spillare, E. A., Hagiwara, K., Hussain, S. P., Bennett, W. P., Forrester, K., Gerwin, B., Serrano, M., Beach, D. H. and Harris, C. C., Mutations and altered expression of p16^ink4 in human cancer. Proc. nat. Acad. Sci. (wash.), 91, 11045–11049 (1994).
10.1073/pnas.91.23.11045
CAS PubMed Web of Science® Google Scholar
Povey, S., Armour, J., Farndon, P., Haines, J. L., Knowles, M., Olopade, F., Pilz, A., White, J. A. and Kwiatkowski, D. J., Report on the Third International Workshop on chromosome 9. Ann. hum. Genet., 58, 117–250 (1994).
10.1111/j.1469-1809.1994.tb01887.x
CAS Web of Science® Google Scholar
Ruppert, J. M., Tokino, K. and Sidransky, D., Evidence for two bladder-Cancer-Suppressor loci on human chromosome 9. Cancer Res., 53, 5093–5095 (1993).
CAS PubMed Web of Science® Google Scholar
Sambrook, J., Fritsch, E. F. and Maniatis, T., Molecular cloning: a laboratory manual ( 2nd ed.), Cold Spring Harbor Laboratory, New York (1989).
10.1111/j.1095-8312.1996.tb01434.x
Google Scholar
Serrano, M., Hannon, G. J. and Beach, D., A new regulatory motifin cell-Cycle control causing specific inhibition of cyclin D/CDK4. Nature (Lond.), 366, 704–707 (1993).
10.1038/366704a0
CAS PubMed Web of Science® Google Scholar
Spruck III, C. H., Gonzales-Zulueta, M., Shibata, A., Simoneau, A. R., Lin, M. F., Gonzales, F., Tsai, Y. C. and Jones, P. A., p16 gene in uncultured tumours. Nature (Lond.), 370, 183–184 (1994).
10.1038/370183a0
PubMed Web of Science® Google Scholar
Stadler, W. M., Sherman, J., Bohlander, S. K., Roulston, D., Dreyling, M., Rukstalis, D. and Olopade, O. I., Homozygous deletions within chromosomal bands 9p21-22 in bladder cancer. Cancer Res., 54, 2060–2063 (1994).
CAS PubMed Google Scholar
Warburton, D., Gersen, S., Yu, M. T., Jackson, C., Handelin, B. and Housman, D., Monochromosomal rodent-Human hybrids from microcell fusion of human lymphoblastoid cells containing an inserted dominant selectable marker. Genomics, 6, 358–366 (1990).
10.1016/0888-7543(90)90577-H
CAS PubMed Web of Science® Google Scholar

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Volume65, Issue6

15 March 1996

Pages 840-846

Growth arrest and suppression of tumorigenicity of bladder carcinoma cell lines induced by the P16/CDKN2 (p16^INK4A, MTS₁) gene and other loci on human chromosome 9

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Growth arrest and suppression of tumorigenicity of bladder carcinoma cell lines induced by the P16/CDKN2 (p16INK4A, MTS1) gene and other loci on human chromosome 9

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Growth arrest and suppression of tumorigenicity of bladder carcinoma cell lines induced by the P16/CDKN2 (p16^INK4A, MTS₁) gene and other loci on human chromosome 9