Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1994 Apr;69(4):680–686. doi: 10.1038/bjc.1994.130

Increased mdr1 gene transcript levels in high-grade carcinoma of the bladder determined by quantitative PCR-based assay.

S C Clifford 1, D J Thomas 1, D E Neal 1, J Lunec 1
PMCID: PMC1968824  PMID: 7908215

Abstract

Overexpression of the multidrug resistance (mdr1) gene has been implicated in resistance to a number of the chemotherapeutic agents currently used in the treatment of bladder cancer (doxorubicin, vincristine and epirubicin). We report the development and validation of a quantitative assay for the determination of mdr1 gene transcript levels based on reverse transcription and the polymerase chain reaction (PCR), sensitive to less than 2-fold variations in transcript levels. Using these techniques, mdr1 mRNA levels were investigated in 32 primary untreated transitional cell carcinomas of the bladder. mdr1 mRNA was detected in all samples, with levels varying between individual tumours over a 63-fold range. These variations were not associated with the proliferative status of the tumour. mdr1 mRNA levels were significantly higher in poorly differentiated high-grade (G3) tumours than in well- and moderately differentiated low-grade (G1 and G2) tumours (P = 0.0057). The results suggest that this relationship may extend to mdr1 mRNA levels being an indicator of poor prognosis, as anticipated on the basis of the observed relationship to tumour stage and grade. No evidence was found to implicate mdr1 mRNA levels as a predictor of tumour recurrence or progression. Given that mdr1 mRNA levels are increased in a proportion of high-grade bladder tumours that are routinely subjected to chemotherapy, we discuss the possibility that mdr1 mRNA levels may be clinically significant as determinants of chemotherapeutic response and outcome in bladder cancer.

Full text

PDF
680

Images in this article

682Figure 1
on p.682

Selected References

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

  1. Becker-André M., Hahlbrock K. Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel approach by a PCR aided transcript titration assay (PATTY). Nucleic Acids Res. 1989 Nov 25;17(22):9437–9446. doi: 10.1093/nar/17.22.9437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benson M. C., Giella J., Whang I. S., Buttyan R., Hensle T. W., Karp F., Olsson C. A. Flow cytometric determination of the multidrug resistant phenotype in transitional cell cancer of the bladder: implications and applications. J Urol. 1991 Oct;146(4):982–987. doi: 10.1016/s0022-5347(17)37981-8. [DOI] [PubMed] [Google Scholar]
  3. Campos L., Guyotat D., Archimbaud E., Calmard-Oriol P., Tsuruo T., Troncy J., Treille D., Fiere D. Clinical significance of multidrug resistance P-glycoprotein expression on acute nonlymphoblastic leukemia cells at diagnosis. Blood. 1992 Jan 15;79(2):473–476. [PubMed] [Google Scholar]
  4. Cattoretti G., Becker M. H., Key G., Duchrow M., Schlüter C., Galle J., Gerdes J. Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave-processed formalin-fixed paraffin sections. J Pathol. 1992 Dec;168(4):357–363. doi: 10.1002/path.1711680404. [DOI] [PubMed] [Google Scholar]
  5. Chan H. S., Bradley G., Thorner P., Haddad G., Gallie B. L., Ling V. A sensitive method for immunocytochemical detection of P-glycoprotein in multidrug-resistant human ovarian carcinoma cell lines. Lab Invest. 1988 Dec;59(6):870–875. [PubMed] [Google Scholar]
  6. Chan H. S., Haddad G., Thorner P. S., DeBoer G., Lin Y. P., Ondrusek N., Yeger H., Ling V. P-glycoprotein expression as a predictor of the outcome of therapy for neuroblastoma. N Engl J Med. 1991 Dec 5;325(23):1608–1614. doi: 10.1056/NEJM199112053252304. [DOI] [PubMed] [Google Scholar]
  7. Chan H. S., Thorner P. S., Haddad G., Ling V. Immunohistochemical detection of P-glycoprotein: prognostic correlation in soft tissue sarcoma of childhood. J Clin Oncol. 1990 Apr;8(4):689–704. doi: 10.1200/JCO.1990.8.4.689. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Endicott J. A., Ling V. The biochemistry of P-glycoprotein-mediated multidrug resistance. Annu Rev Biochem. 1989;58:137–171. doi: 10.1146/annurev.bi.58.070189.001033. [DOI] [PubMed] [Google Scholar]
  10. Fojo A. T., Ueda K., Slamon D. J., Poplack D. G., Gottesman M. M., Pastan I. Expression of a multidrug-resistance gene in human tumors and tissues. Proc Natl Acad Sci U S A. 1987 Jan;84(1):265–269. doi: 10.1073/pnas.84.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldstein L. J., Galski H., Fojo A., Willingham M., Lai S. L., Gazdar A., Pirker R., Green A., Crist W., Brodeur G. M. Expression of a multidrug resistance gene in human cancers. J Natl Cancer Inst. 1989 Jan 18;81(2):116–124. doi: 10.1093/jnci/81.2.116. [DOI] [PubMed] [Google Scholar]
  12. Gonzalez I. L., Schmickel R. D. The human 18S ribosomal RNA gene: evolution and stability. Am J Hum Genet. 1986 Apr;38(4):419–427. [PMC free article] [PubMed] [Google Scholar]
  13. Harker W. G., Meyers F. J., Freiha F. S., Palmer J. M., Shortliffe L. D., Hannigan J. F., McWhirter K. M., Torti F. M. Cisplatin, methotrexate, and vinblastine (CMV): an effective chemotherapy regimen for metastatic transitional cell carcinoma of the urinary tract. A Northern California Oncology Group study. J Clin Oncol. 1985 Nov;3(11):1463–1470. doi: 10.1200/JCO.1985.3.11.1463. [DOI] [PubMed] [Google Scholar]
  14. Hirsch C. A. Quantitative determination of the ribosomal ribonucleic acid content of liver and Novikoff hepatoma from fed and from fasted rats. J Biol Chem. 1967 Jun 25;242(12):2822–2827. [PubMed] [Google Scholar]
  15. Johnson L. F., Abelson H. T., Penman S., Green H. The relative amounts of the cytoplasmic RNA species in normal, transformed and senescent cultured cell lines. J Cell Physiol. 1977 Mar;90(3):465–470. doi: 10.1002/jcp.1040900310. [DOI] [PubMed] [Google Scholar]
  16. Kiemeney L. A., Witjes J. A., Verbeek A. L., Heijbroek R. P., Debruyne F. M. The clinical epidemiology of superficial bladder cancer. Dutch South-East Cooperative Urological Group. Br J Cancer. 1993 Apr;67(4):806–812. doi: 10.1038/bjc.1993.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kimiya K., Naito S., Soejima T., Sakamoto N., Kotoh S., Kumazawa J., Tsuruo T. Establishment and characterization of doxorubicin-resistant human bladder cancer cell line, KK47/ADM. J Urol. 1992 Aug;148(2 Pt 1):441–445. doi: 10.1016/s0022-5347(17)36624-7. [DOI] [PubMed] [Google Scholar]
  18. Kuwazuru Y., Yoshimura A., Hanada S., Utsunomiya A., Makino T., Ishibashi K., Kodama M., Iwahashi M., Arima T., Akiyama S. Expression of the multidrug transporter, P-glycoprotein, in acute leukemia cells and correlation to clinical drug resistance. Cancer. 1990 Sep 1;66(5):868–873. doi: 10.1002/1097-0142(19900901)66:5<868::aid-cncr2820660510>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  19. Lowe T., Sharefkin J., Yang S. Q., Dieffenbach C. W. A computer program for selection of oligonucleotide primers for polymerase chain reactions. Nucleic Acids Res. 1990 Apr 11;18(7):1757–1761. doi: 10.1093/nar/18.7.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Naito S., Sakamoto N., Kotoh S., Goto K., Matsumoto T., Kumazawa J. Correlation between the expression of P-glycoprotein and multidrug-resistant phenotype in transitional cell carcinoma of the urinary tract. Eur Urol. 1992;22(2):158–162. doi: 10.1159/000474745. [DOI] [PubMed] [Google Scholar]
  21. Noonan K. E., Beck C., Holzmayer T. A., Chin J. E., Wunder J. S., Andrulis I. L., Gazdar A. F., Willman C. L., Griffith B., Von Hoff D. D. Quantitative analysis of MDR1 (multidrug resistance) gene expression in human tumors by polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7160–7164. doi: 10.1073/pnas.87.18.7160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Peto R., Pike M. C., Armitage P., Breslow N. E., Cox D. R., Howard S. V., Mantel N., McPherson K., Peto J., Smith P. G. Design and analysis of randomized clinical trials requiring prolonged observation of each patient. II. analysis and examples. Br J Cancer. 1977 Jan;35(1):1–39. doi: 10.1038/bjc.1977.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pirker R., Wallner J., Geissler K., Linkesch W., Haas O. A., Bettelheim P., Hopfner M., Scherrer R., Valent P., Havelec L. MDR1 gene expression and treatment outcome in acute myeloid leukemia. J Natl Cancer Inst. 1991 May 15;83(10):708–712. doi: 10.1093/jnci/83.10.708. [DOI] [PubMed] [Google Scholar]
  24. Shen D. W., Fojo A., Roninson I. B., Chin J. E., Soffir R., Pastan I., Gottesman M. M. Multidrug resistance of DNA-mediated transformants is linked to transfer of the human mdr1 gene. Mol Cell Biol. 1986 Nov;6(11):4039–4045. doi: 10.1128/mcb.6.11.4039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Twentyman P. R., Fox N. E., Wright K. A., Bleehen N. M. Derivation and preliminary characterisation of adriamycin resistant lines of human lung cancer cells. Br J Cancer. 1986 Apr;53(4):529–537. doi: 10.1038/bjc.1986.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Verrelle P., Meissonnier F., Fonck Y., Feillel V., Dionet C., Kwiatkowski F., Plagne R., Chassagne J. Clinical relevance of immunohistochemical detection of multidrug resistance P-glycoprotein in breast carcinoma. J Natl Cancer Inst. 1991 Jan 16;83(2):111–116. doi: 10.1093/jnci/83.2.111. [DOI] [PubMed] [Google Scholar]
  27. Wang A. M., Doyle M. V., Mark D. F. Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9717–9721. doi: 10.1073/pnas.86.24.9717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zastawny R. L., Salvino R., Chen J., Benchimol S., Ling V. The core promoter region of the P-glycoprotein gene is sufficient to confer differential responsiveness to wild-type and mutant p53. Oncogene. 1993 Jun;8(6):1529–1535. [PubMed] [Google Scholar]
  29. van der Bliek A. M., Borst P. Multidrug resistance. Adv Cancer Res. 1989;52:165–203. doi: 10.1016/s0065-230x(08)60213-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES