Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1991 Apr;63(4):601–608. doi: 10.1038/bjc.1991.139

Amplification and over-expression of c-erbB-2 in transitional cell carcinoma of the urinary bladder.

L M Coombs 1, D A Pigott 1, E Sweeney 1, A J Proctor 1, M E Eydmann 1, C Parkinson 1, M A Knowles 1
PMCID: PMC1972370  PMID: 1673627

Abstract

The structure and expression of the proto-oncogene c-erbB-2 was studied in 86 patients with transitional cell carcinoma. Initial tissue samples comprised 37 grade 1, 32 grade 2 and 13 grade 3 tumours and four cases of carcinoma in situ. At the time of this first tumour sample, amplification of the c-erbB-2 gene was demonstrated by Southern blotting in 1/37 grade 1, 5/32 grade 2 and 6/13 grade 3 tumours (0.005 less than P less than 0.01). Tumour 're-occurrences' were obtained from 23 of these patients on one or more occasions. Amplification was detected in re-occurrences from seven of these 23, none of whom showed amplification in the first tumour sample. DNA was also extracted from exfoliated cells in urine collected from five cases of carcinoma in situ and c-erbB-2 amplification was demonstrated in one of these. No gene amplification was identified in patients' lymphocytes, ten biopsies of normal urothelium and 22 various intravesical pathologies. Increased expression of c-erbB-2 mRNA correlated with amplification of the gene. In addition, raised levels of mRNA were seen in the absence of gene amplification in six tumours. Immunoblotting using the polyclonal antibody 21N, raised against the c-terminus of the c-erbB-2 protein demonstrated increased amounts of a 185 kD immunoreactive protein in tumours with increased c-erbB-2 gene copy number compared with control tissues. In some tumours with high c-erbB-2 gene copy number, a 155 kD immunoreactive protein not detected in controls was expressed at higher level than the 185 kD protein. Immunocytochemistry using a monoclonal antibody AB-3, raised against the c-terminus of the c-erbB-2 protein, showed a positive reaction in the cytoplasm and cell membrane of tumours with gene amplification and in 40% of tumours with no amplification. An association was found between c-erbB-2 amplification and over-expression and the development of tumour re-occurrences. We suggest that c-erbB-2 amplification and over-expression may provide a useful molecular marker in transitional cell carcinoma of the bladder and merits further investigation as a potential prognostic indicator.

Full text

PDF
604

Images in this article

604Figure 1
on p.604
604Figure 2
on p.604
605Figure 4
on p.605
605Figure 3
on p.605

Selected References

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

  1. Abel P. D., Henderson D., Bennett M. K., Hall R. R., Williams G. Differing interpretations by pathologists of the pT category and grade of transitional cell cancer of the bladder. Br J Urol. 1988 Oct;62(4):339–342. doi: 10.1111/j.1464-410x.1988.tb04361.x. [DOI] [PubMed] [Google Scholar]
  2. Akiyama T., Saito T., Ogawara H., Toyoshima K., Yamamoto T. Tumor promoter and epidermal growth factor stimulate phosphorylation of the c-erbB-2 gene product in MKN-7 human adenocarcinoma cells. Mol Cell Biol. 1988 Mar;8(3):1019–1026. doi: 10.1128/mcb.8.3.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Akiyama T., Sudo C., Ogawara H., Toyoshima K., Yamamoto T. The product of the human c-erbB-2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activity. Science. 1986 Jun 27;232(4758):1644–1646. doi: 10.1126/science.3012781. [DOI] [PubMed] [Google Scholar]
  4. Bargmann C. I., Hung M. C., Weinberg R. A. Multiple independent activations of the neu oncogene by a point mutation altering the transmembrane domain of p185. Cell. 1986 Jun 6;45(5):649–657. doi: 10.1016/0092-8674(86)90779-8. [DOI] [PubMed] [Google Scholar]
  5. Bargmann C. I., Hung M. C., Weinberg R. A. The neu oncogene encodes an epidermal growth factor receptor-related protein. Nature. 1986 Jan 16;319(6050):226–230. doi: 10.1038/319226a0. [DOI] [PubMed] [Google Scholar]
  6. Benchimol S., Lamb P., Crawford L. V., Sheer D., Shows T. B., Bruns G. A., Peacock J. Transformation associated p53 protein is encoded by a gene on human chromosome 17. Somat Cell Mol Genet. 1985 Sep;11(5):505–510. doi: 10.1007/BF01534845. [DOI] [PubMed] [Google Scholar]
  7. Berger M. S., Greenfield C., Gullick W. J., Haley J., Downward J., Neal D. E., Harris A. L., Waterfield M. D. Evaluation of epidermal growth factor receptors in bladder tumours. Br J Cancer. 1987 Nov;56(5):533–537. doi: 10.1038/bjc.1987.238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Berger M. S., Locher G. W., Saurer S., Gullick W. J., Waterfield M. D., Groner B., Hynes N. E. Correlation of c-erbB-2 gene amplification and protein expression in human breast carcinoma with nodal status and nuclear grading. Cancer Res. 1988 Mar 1;48(5):1238–1243. [PubMed] [Google Scholar]
  9. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  10. Coombs L. M., Pigott D., Proctor A., Eydmann M., Denner J., Knowles M. A. Simultaneous isolation of DNA, RNA, and antigenic protein exhibiting kinase activity from small tumor samples using guanidine isothiocyanate. Anal Biochem. 1990 Aug 1;188(2):338–343. doi: 10.1016/0003-2697(90)90617-i. [DOI] [PubMed] [Google Scholar]
  11. Coussens L., Yang-Feng T. L., Liao Y. C., Chen E., Gray A., McGrath J., Seeburg P. H., Libermann T. A., Schlessinger J., Francke U. Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science. 1985 Dec 6;230(4730):1132–1139. doi: 10.1126/science.2999974. [DOI] [PubMed] [Google Scholar]
  12. D'Emilia J., Bulovas K., D'Ercole K., Wolf B., Steele G., Jr, Summerhayes I. C. Expression of the c-erbB-2 gene product (p185) at different stages of neoplastic progression in the colon. Oncogene. 1989 Oct;4(10):1233–1239. [PubMed] [Google Scholar]
  13. De Potter C. R., Quatacker J., Maertens G., Van Daele S., Pauwels C., Verhofstede C., Eechaute W., Roels H. The subcellular localization of the neu protein in human normal and neoplastic cells. Int J Cancer. 1989 Dec 15;44(6):969–974. doi: 10.1002/ijc.2910440604. [DOI] [PubMed] [Google Scholar]
  14. Farber E. The multistep nature of cancer development. Cancer Res. 1984 Oct;44(10):4217–4223. [PubMed] [Google Scholar]
  15. Fearon E. R., Feinberg A. P., Hamilton S. H., Vogelstein B. Loss of genes on the short arm of chromosome 11 in bladder cancer. 1985 Nov 28-Dec 4Nature. 318(6044):377–380. doi: 10.1038/318377a0. [DOI] [PubMed] [Google Scholar]
  16. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  17. Fort P., Marty L., Piechaczyk M., el Sabrouty S., Dani C., Jeanteur P., Blanchard J. M. Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res. 1985 Mar 11;13(5):1431–1442. doi: 10.1093/nar/13.5.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fujita J., Srivastava S. K., Kraus M. H., Rhim J. S., Tronick S. R., Aaronson S. A. Frequency of molecular alterations affecting ras protooncogenes in human urinary tract tumors. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3849–3853. doi: 10.1073/pnas.82.11.3849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gullick W. J., Berger M. S., Bennett P. L., Rothbard J. B., Waterfield M. D. Expression of the c-erbB-2 protein in normal and transformed cells. Int J Cancer. 1987 Aug 15;40(2):246–254. doi: 10.1002/ijc.2910400221. [DOI] [PubMed] [Google Scholar]
  20. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  21. King C. R., Kraus M. H., Aaronson S. A. Amplification of a novel v-erbB-related gene in a human mammary carcinoma. Science. 1985 Sep 6;229(4717):974–976. doi: 10.1126/science.2992089. [DOI] [PubMed] [Google Scholar]
  22. Kokai Y., Myers J. N., Wada T., Brown V. I., LeVea C. M., Davis J. G., Dobashi K., Greene M. I. Synergistic interaction of p185c-neu and the EGF receptor leads to transformation of rodent fibroblasts. Cell. 1989 Jul 28;58(2):287–292. doi: 10.1016/0092-8674(89)90843-x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Lau Y. F., Kan Y. W. Direct isolation of the functional human thymidine kinase gene with a cosmid shuttle vector. Proc Natl Acad Sci U S A. 1984 Jan;81(2):414–418. doi: 10.1073/pnas.81.2.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lemoine N. R., Staddon S., Dickson C., Barnes D. M., Gullick W. J. Absence of activating transmembrane mutations in the c-erbB-2 proto-oncogene in human breast cancer. Oncogene. 1990 Feb;5(2):237–239. [PubMed] [Google Scholar]
  26. Neal D. E., Marsh C., Bennett M. K., Abel P. D., Hall R. R., Sainsbury J. R., Harris A. L. Epidermal-growth-factor receptors in human bladder cancer: comparison of invasive and superficial tumours. Lancet. 1985 Feb 16;1(8425):366–368. doi: 10.1016/s0140-6736(85)91386-8. [DOI] [PubMed] [Google Scholar]
  27. Ooms E. C., Anderson W. A., Alons C. L., Boon M. E., Veldhuizen R. W. Analysis of the performance of pathologists in the grading of bladder tumors. Hum Pathol. 1983 Feb;14(2):140–143. doi: 10.1016/s0046-8177(83)80242-1. [DOI] [PubMed] [Google Scholar]
  28. Pryor J. P. Factors influencing the survival of patients with transitional cell tumours of the urinary bladder. Br J Urol. 1973 Dec;45(6):586–592. doi: 10.1111/j.1464-410x.1973.tb12227.x. [DOI] [PubMed] [Google Scholar]
  29. Raghavan D., Shipley W. U., Garnick M. B., Russell P. J., Richie J. P. Biology and management of bladder cancer. N Engl J Med. 1990 Apr 19;322(16):1129–1138. doi: 10.1056/NEJM199004193221607. [DOI] [PubMed] [Google Scholar]
  30. Schechter A. L., Hung M. C., Vaidyanathan L., Weinberg R. A., Yang-Feng T. L., Francke U., Ullrich A., Coussens L. The neu gene: an erbB-homologous gene distinct from and unlinked to the gene encoding the EGF receptor. Science. 1985 Sep 6;229(4717):976–978. doi: 10.1126/science.2992090. [DOI] [PubMed] [Google Scholar]
  31. Schechter A. L., Stern D. F., Vaidyanathan L., Decker S. J., Drebin J. A., Greene M. I., Weinberg R. A. The neu oncogene: an erb-B-related gene encoding a 185,000-Mr tumour antigen. Nature. 1984 Dec 6;312(5994):513–516. doi: 10.1038/312513a0. [DOI] [PubMed] [Google Scholar]
  32. Semba K., Kamata N., Toyoshima K., Yamamoto T. A v-erbB-related protooncogene, c-erbB-2, is distinct from the c-erbB-1/epidermal growth factor-receptor gene and is amplified in a human salivary gland adenocarcinoma. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6497–6501. doi: 10.1073/pnas.82.19.6497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shih C., Padhy L. C., Murray M., Weinberg R. A. Transforming genes of carcinomas and neuroblastomas introduced into mouse fibroblasts. Nature. 1981 Mar 19;290(5803):261–264. doi: 10.1038/290261a0. [DOI] [PubMed] [Google Scholar]
  34. Slamon D. J., Godolphin W., Jones L. A., Holt J. A., Wong S. G., Keith D. E., Levin W. J., Stuart S. G., Udove J., Ullrich A. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science. 1989 May 12;244(4905):707–712. doi: 10.1126/science.2470152. [DOI] [PubMed] [Google Scholar]
  35. Smith K., Fennelly J. A., Neal D. E., Hall R. R., Harris A. L. Characterization and quantitation of the epidermal growth factor receptor in invasive and superficial bladder tumors. Cancer Res. 1989 Nov 1;49(21):5810–5815. [PubMed] [Google Scholar]
  36. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  37. Tavassoli M., Quirke P., Farzaneh F., Lock N. J., Mayne L. V., Kirkham N. c-erbB-2/c-erbA co-amplification indicative of lymph node metastasis, and c-myc amplification of high tumour grade, in human breast carcinoma. Br J Cancer. 1989 Oct;60(4):505–510. doi: 10.1038/bjc.1989.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Visvanathan K. V., Pocock R. D., Summerhayes I. C. Preferential and novel activation of H-ras in human bladder carcinomas. Oncogene Res. 1988;3(1):77–86. [PubMed] [Google Scholar]
  41. Wright C., Mellon K., Neal D. E., Johnston P., Corbett I. P., Horne C. H. Expression of c-erbB-2 protein product in bladder cancer. Br J Cancer. 1990 Nov;62(5):764–765. doi: 10.1038/bjc.1990.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yamamoto T., Ikawa S., Akiyama T., Semba K., Nomura N., Miyajima N., Saito T., Toyoshima K. Similarity of protein encoded by the human c-erb-B-2 gene to epidermal growth factor receptor. Nature. 1986 Jan 16;319(6050):230–234. doi: 10.1038/319230a0. [DOI] [PubMed] [Google Scholar]
  43. Yokota J., Yamamoto T., Toyoshima K., Terada M., Sugimura T., Battifora H., Cline M. J. Amplification of c-erbB-2 oncogene in human adenocarcinomas in vivo. Lancet. 1986 Apr 5;1(8484):765–767. doi: 10.1016/s0140-6736(86)91782-4. [DOI] [PubMed] [Google Scholar]
  44. Zhou D. J., Ahuja H., Cline M. J. Proto-oncogene abnormalities in human breast cancer: c-ERBB-2 amplification does not correlate with recurrence of disease. Oncogene. 1989 Jan;4(1):105–108. [PubMed] [Google Scholar]
  45. van Tuinen P., Rich D. C., Summers K. M., Ledbetter D. H. Regional mapping panel for human chromosome 17: application to neurofibromatosis type 1. Genomics. 1987 Dec;1(4):374–381. doi: 10.1016/0888-7543(87)90042-5. [DOI] [PubMed] [Google Scholar]
  46. van de Vijver M. J., Peterse J. L., Mooi W. J., Wisman P., Lomans J., Dalesio O., Nusse R. Neu-protein overexpression in breast cancer. Association with comedo-type ductal carcinoma in situ and limited prognostic value in stage II breast cancer. N Engl J Med. 1988 Nov 10;319(19):1239–1245. doi: 10.1056/NEJM198811103191902. [DOI] [PubMed] [Google Scholar]
  47. van de Vijver M., van de Bersselaar R., Devilee P., Cornelisse C., Peterse J., Nusse R. Amplification of the neu (c-erbB-2) oncogene in human mammmary tumors is relatively frequent and is often accompanied by amplification of the linked c-erbA oncogene. Mol Cell Biol. 1987 May;7(5):2019–2023. doi: 10.1128/mcb.7.5.2019. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES