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. 1998 Feb;77(4):556–561. doi: 10.1038/bjc.1998.90

Role of glutathione, glutathione S-transferases and multidrug resistance-related proteins in cisplatin sensitivity of head and neck cancer cell lines.

M J Welters 1, A M Fichtinger-Schepman 1, R A Baan 1, M J Flens 1, R J Scheper 1, B J Braakhuis 1
PMCID: PMC2149938  PMID: 9484811

Abstract

Resistance to chemotherapy is a major problem in the treatment of patients with head and neck squamous cell carcinoma (HNSCC). Important factors involved are drug detoxification by glutathione (GSH) and reduced drug accumulation due to active transport out of the cell by so-called 'multidrug resistance-related proteins'. We have studied a panel of eight HNSCC cell lines showing differences in sensitivity to the anti-cancer drug cisplatin. Our previous studies indicated that the IC50 values were inversely correlated with the intracellular accumulation of platinum (Pt). In the present study, cellular GSH levels were found not to be related to the IC50 values. The expression levels of the enzymes glutathione S-transferase (GST) alpha, mu, and pi, the multidrug resistance-related proteins P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP) and the lung resistance protein (LRP) were determined semiquantitatively by means of immunocytochemistry. The levels of the GSTs, P-gp and LRP were not found to be correlated with the IC50 values of the HNSCC cell lines. Surprisingly, however, an inverse correlation was found between MRP levels and IC50 values. The MRP expression levels were in agreement with the results of the MRP functional assay, based on the transport of calcein across the cell membrane as performed for two of the cell lines. Further studies should prove whether other pump mechanisms or DNA repair are involved in the cisplatin accumulation and the subsequent HNSCC cell growth inhibition.

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Selected References

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  1. Bernal S. D., Speak J. A., Boeheim K., Dreyfuss A. I., Wright J. E., Teicher B. A., Rosowsky A., Tsao S. W., Wong Y. C. Reduced membrane protein associated with resistance of human squamous carcinoma cells to methotrexate and cis-platinum. Mol Cell Biochem. 1990 Jun 1;95(1):61–70. doi: 10.1007/BF00219531. [DOI] [PubMed] [Google Scholar]
  2. Biedler J. L. Genetic aspects of multidrug resistance. Cancer. 1992 Sep 15;70(6 Suppl):1799–1809. doi: 10.1002/1097-0142(19920915)70:4+<1799::aid-cncr2820701623>3.0.co;2-b. [DOI] [PubMed] [Google Scholar]
  3. Bongers V., Snow G. B., de Vries N., Cattan A. R., Hall A. G., van der Waal I., Braakhuis B. J. Second primary head and neck squamous cell carcinoma predicted by the glutathione S-transferase expression in healthy tissue in the direct vicinity of the first tumor. Lab Invest. 1995 Oct;73(4):503–510. [PubMed] [Google Scholar]
  4. Braakhuis B. J., Jansen G., Noordhuis P., Kegel A., Peters G. J. Importance of pharmacodynamics in the in vitro antiproliferative activity of the antifolates methotrexate and 10-ethyl-10-deazaaminopterin against human head and neck squamous cell carcinoma. Biochem Pharmacol. 1993 Dec 14;46(12):2155–2161. doi: 10.1016/0006-2952(93)90604-u. [DOI] [PubMed] [Google Scholar]
  5. Brock I., Hipfner D. R., Nielsen B. S., Jensen P. B., Deeley R. G., Cole S. P., Sehested M. Sequential coexpression of the multidrug resistance genes MRP and mdr1 and their products in VP-16 (etoposide)-selected H69 small cell lung cancer cells. Cancer Res. 1995 Feb 1;55(3):459–462. [PubMed] [Google Scholar]
  6. Broxterman H. J., Giaccone G., Lankelma J. Multidrug resistance proteins and other drug transport-related resistance to natural product agents. Curr Opin Oncol. 1995 Nov;7(6):532–540. doi: 10.1097/00001622-199511000-00011. [DOI] [PubMed] [Google Scholar]
  7. Broxterman H. J., Lankelma J., Pinedo H. M. How to probe clinical tumour samples for P-glycoprotein and multidrug resistance-associated protein. Eur J Cancer. 1996 Jun;32A(6):1024–1033. doi: 10.1016/0959-8049(96)00045-7. [DOI] [PubMed] [Google Scholar]
  8. Feller N., Broxterman H. J., Währer D. C., Pinedo H. M. ATP-dependent efflux of calcein by the multidrug resistance protein (MRP): no inhibition by intracellular glutathione depletion. FEBS Lett. 1995 Jul 17;368(2):385–388. doi: 10.1016/0014-5793(95)00677-2. [DOI] [PubMed] [Google Scholar]
  9. Flens M. J., Izquierdo M. A., Scheffer G. L., Fritz J. M., Meijer C. J., Scheper R. J., Zaman G. J. Immunochemical detection of the multidrug resistance-associated protein MRP in human multidrug-resistant tumor cells by monoclonal antibodies. Cancer Res. 1994 Sep 1;54(17):4557–4563. [PubMed] [Google Scholar]
  10. Goto S., Yoshida K., Morikawa T., Urata Y., Suzuki K., Kondo T. Augmentation of transport for cisplatin-glutathione adduct in cisplatin-resistant cancer cells. Cancer Res. 1995 Oct 1;55(19):4297–4301. [PubMed] [Google Scholar]
  11. Hayes J. D., Wolf C. R. Molecular mechanisms of drug resistance. Biochem J. 1990 Dec 1;272(2):281–295. doi: 10.1042/bj2720281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hermsen M. A., Joenje H., Arwert F., Welters M. J., Braakhuis B. J., Bagnay M., Westerveld A., Slater R. Centromeric breakage as a major cause of cytogenetic abnormalities in oral squamous cell carcinoma. Genes Chromosomes Cancer. 1996 Jan;15(1):1–9. doi: 10.1002/(SICI)1098-2264(199601)15:1<1::AID-GCC1>3.0.CO;2-8. [DOI] [PubMed] [Google Scholar]
  13. Ishikawa T., Ali-Osman F. Glutathione-associated cis-diamminedichloroplatinum(II) metabolism and ATP-dependent efflux from leukemia cells. Molecular characterization of glutathione-platinum complex and its biological significance. J Biol Chem. 1993 Sep 25;268(27):20116–20125. [PubMed] [Google Scholar]
  14. Ishikawa T., Bao J. J., Yamane Y., Akimaru K., Frindrich K., Wright C. D., Kuo M. T. Coordinated induction of MRP/GS-X pump and gamma-glutamylcysteine synthetase by heavy metals in human leukemia cells. J Biol Chem. 1996 Jun 21;271(25):14981–14988. doi: 10.1074/jbc.271.25.14981. [DOI] [PubMed] [Google Scholar]
  15. Ishikawa T., Wright C. D., Ishizuka H. GS-X pump is functionally overexpressed in cis-diamminedichloroplatinum (II)-resistant human leukemia HL-60 cells and down-regulated by cell differentiation. J Biol Chem. 1994 Nov 18;269(46):29085–29093. [PubMed] [Google Scholar]
  16. Izquierdo M. A., Scheffer G. L., Flens M. J., Giaccone G., Broxterman H. J., Meijer C. J., van der Valk P., Scheper R. J. Broad distribution of the multidrug resistance-related vault lung resistance protein in normal human tissues and tumors. Am J Pathol. 1996 Mar;148(3):877–887. [PMC free article] [PubMed] [Google Scholar]
  17. Izquierdo M. A., van der Zee A. G., Vermorken J. B., van der Valk P., Beliën J. A., Giaccone G., Scheffer G. L., Flens M. J., Pinedo H. M., Kenemans P. Drug resistance-associated marker Lrp for prediction of response to chemotherapy and prognoses in advanced ovarian carcinoma. J Natl Cancer Inst. 1995 Aug 16;87(16):1230–1237. doi: 10.1093/jnci/87.16.1230. [DOI] [PubMed] [Google Scholar]
  18. Karlsson M., Jungnelius U., Aamdal S., Boeryd B., Carstensen J., Kågedal B., Westberg R., Wingren S. Correlation of DNA ploidy and S-phase fraction with chemotherapeutic response and survival in a randomized study of disseminated malignant melanoma. Int J Cancer. 1996 Jan 3;65(1):1–5. doi: 10.1002/(SICI)1097-0215(19960103)65:1<1::AID-IJC1>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  19. Kelley D. J., Pavelic Z. P., Gapany M., Stambrook P., Pavelic L., Gapany S., Gluckman J. L. Detection of P-glycoprotein in squamous cell carcinomas of the head and neck. Arch Otolaryngol Head Neck Surg. 1993 Apr;119(4):411–414. doi: 10.1001/archotol.1993.01880160057009. [DOI] [PubMed] [Google Scholar]
  20. Kramer R. A., Zakher J., Kim G. Role of the glutathione redox cycle in acquired and de novo multidrug resistance. Science. 1988 Aug 5;241(4866):694–697. doi: 10.1126/science.3399900. [DOI] [PubMed] [Google Scholar]
  21. Kurokawa H., Ishida T., Nishio K., Arioka H., Sata M., Fukumoto H., Miura M., Saijo N. Gamma-glutamylcysteine synthetase gene overexpression results in increased activity of the ATP-dependent glutathione S-conjugate export pump and cisplatin resistance. Biochem Biophys Res Commun. 1995 Nov 2;216(1):258–264. doi: 10.1006/bbrc.1995.2618. [DOI] [PubMed] [Google Scholar]
  22. Loe D. W., Deeley R. G., Cole S. P. Biology of the multidrug resistance-associated protein, MRP. Eur J Cancer. 1996 Jun;32A(6):945–957. doi: 10.1016/0959-8049(96)00046-9. [DOI] [PubMed] [Google Scholar]
  23. Meijer C., Mulder N. H., Timmer-Bosscha H., Sluiter W. J., Meersma G. J., de Vries E. G. Relationship of cellular glutathione to the cytotoxicity and resistance of seven platinum compounds. Cancer Res. 1992 Dec 15;52(24):6885–6889. [PubMed] [Google Scholar]
  24. Mistry P., Kelland L. R., Abel G., Sidhar S., Harrap K. R. The relationships between glutathione, glutathione-S-transferase and cytotoxicity of platinum drugs and melphalan in eight human ovarian carcinoma cell lines. Br J Cancer. 1991 Aug;64(2):215–220. doi: 10.1038/bjc.1991.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Müller M., Meijer C., Zaman G. J., Borst P., Scheper R. J., Mulder N. H., de Vries E. G., Jansen P. L. Overexpression of the gene encoding the multidrug resistance-associated protein results in increased ATP-dependent glutathione S-conjugate transport. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):13033–13037. doi: 10.1073/pnas.91.26.13033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Neuschwander-Tetri B. A., Roll F. J. Glutathione measurement by high-performance liquid chromatography separation and fluorometric detection of the glutathione-orthophthalaldehyde adduct. Anal Biochem. 1989 Jun;179(2):236–241. doi: 10.1016/0003-2697(89)90121-8. [DOI] [PubMed] [Google Scholar]
  27. Nooter K., Westerman A. M., Flens M. J., Zaman G. J., Scheper R. J., van Wingerden K. E., Burger H., Oostrum R., Boersma T., Sonneveld P. Expression of the multidrug resistance-associated protein (MRP) gene in human cancers. Clin Cancer Res. 1995 Nov;1(11):1301–1310. [PubMed] [Google Scholar]
  28. O'Brien M. L., Tew K. D. Glutathione and related enzymes in multidrug resistance. Eur J Cancer. 1996 Jun;32A(6):967–978. doi: 10.1016/0959-8049(96)00051-2. [DOI] [PubMed] [Google Scholar]
  29. Scheffer G. L., Wijngaard P. L., Flens M. J., Izquierdo M. A., Slovak M. L., Pinedo H. M., Meijer C. J., Clevers H. C., Scheper R. J. The drug resistance-related protein LRP is the human major vault protein. Nat Med. 1995 Jun;1(6):578–582. doi: 10.1038/nm0695-578. [DOI] [PubMed] [Google Scholar]
  30. Scheper R. J., Broxterman H. J., Scheffer G. L., Kaaijk P., Dalton W. S., van Heijningen T. H., van Kalken C. K., Slovak M. L., de Vries E. G., van der Valk P. Overexpression of a M(r) 110,000 vesicular protein in non-P-glycoprotein-mediated multidrug resistance. Cancer Res. 1993 Apr 1;53(7):1475–1479. [PubMed] [Google Scholar]
  31. Taniguchi K., Wada M., Kohno K., Nakamura T., Kawabe T., Kawakami M., Kagotani K., Okumura K., Akiyama S., Kuwano M. A human canalicular multispecific organic anion transporter (cMOAT) gene is overexpressed in cisplatin-resistant human cancer cell lines with decreased drug accumulation. Cancer Res. 1996 Sep 15;56(18):4124–4129. [PubMed] [Google Scholar]
  32. Vokes E. E., Weichselbaum R. R., Lippman S. M., Hong W. K. Head and neck cancer. N Engl J Med. 1993 Jan 21;328(3):184–194. doi: 10.1056/NEJM199301213280306. [DOI] [PubMed] [Google Scholar]
  33. Welters M. J., Fichtinger-Schepman A. M., Baan R. A., Hermsen M. A., van der Vijgh W. J., Cloos J., Braakhuis B. J. Relationship between the parameters cellular differentiation, doubling time and platinum accumulation and cisplatin sensitivity in a panel of head and neck cancer cell lines. Int J Cancer. 1997 May 2;71(3):410–415. doi: 10.1002/(sici)1097-0215(19970502)71:3<410::aid-ijc18>3.0.co;2-j. [DOI] [PubMed] [Google Scholar]
  34. Yellin S. A., Davidson B. J., Pinto J. T., Sacks P. G., Qiao C., Schantz S. P. Relationship of glutathione and glutathione-S-transferase to cisplatin sensitivity in human head and neck squamous carcinoma cell lines. Cancer Lett. 1994 Oct 14;85(2):223–232. doi: 10.1016/0304-3835(94)90279-8. [DOI] [PubMed] [Google Scholar]
  35. de Vries E. G., Müller M., Meijer C., Jansen P. L., Mulder N. H. Role of the glutathione S-conjugate pump in cisplatin resistance. J Natl Cancer Inst. 1995 Apr 5;87(7):537–538. doi: 10.1093/jnci/87.7.537. [DOI] [PubMed] [Google Scholar]

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