Role of reactive oxygen species in cis-dichlorodiammineplatinum-induced cytotoxicity on bladder cancer cells

This study was undertaken to investigate the intracellular induction of reactive oxygen species (ROS) by cis-dichlorodiammineplatinum (CDDP) and the augmentation of their cytotoxicity in bladder cancer cells (KU7) by enhancement of ROS generation by the glutathione (GSH) depletors buthionine sulphoximine (BSO) and diethylmaleate (DEM). CDDP-induced cytotoxicity in KU7 cells and its modulation by GSH depletors were determined using spectrophotometric measurement with crystal violet staining. The effects of GSH depletors on intracellular GSH levels were confirmed using the GSH reductase-DTNB recycling method. Intracellular ROS generation induced by CDDP with or without GSH depletors was estimated from the amount of intracellular dichlorofluorescein (DCF), an oxidized product of dichlorofluorescein (DCFH), which was measured with an anchored cell analysis and sorting system. The cytotoxic effects of CDDP (IC50 15.0 +/- 2.5 microM) were significantly enhanced by BSO (IC50 9.3 +/- 2.6 microM, P < 0.01) and DEM (IC50 10.3 +/- 0.3 microM, P <0.01). BSO and DEM produced a significant depletion in intracellular GSH levels (9.6 +/- 0.4 nmol 10(-6) cells, 17.9 +/- 1.0 nmol 10(-6) cells) compared with the controls (30.5 +/- 0.6 nmol 10(-6) cells). Intracellular DCF production in KU7 cells treated with CDDP (1.35 +/- 0.33 microM) was significantly enhanced by the addition of BSO (4.43 +/- 0.33 microM) or DEM (3.12 +/- 0.22 microM) at 150 min. These results suggest that ROS may play a substantial role in CDDP-induced cytotoxicity and that GSH depletors augment its cytotoxicity through an enhancement of ROS generation in bladder cancer cells.

Abstract licence: CC BY

1. 1.

   Anderson M. E., Naganuma A., Meister A. Protection against cisplatin toxicity by administration of glutathione ester. FASEB J. 1990;4(14):3251-3255.

   Not in our library10.1096/fasebj.4.14.2227215 (opens in a new tab)

2. 2.

   Bass D. A., Parce J. W., Dechatelet L. R., Szejda P., Seeds M. C., Thomas M. Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation. J Immunol. 1983;130(4):1910-1917.

   Not in our library

3. 3.

   Bulkley G. B. The role of oxygen free radicals in human disease processes. Surgery. 1983;94(3):407-411.

   Not in our library

4. 4.

   Busciglio J., Yankner B. A. Apoptosis and increased generation of reactive oxygen species in Down's syndrome neurons in vitro. Nature. 1995;378(6559):776-779.

   Not in our library10.1038/378776a0 (opens in a new tab)

5. 5.

   Cathcart R., Schwiers E., Ames B. N. Detection of picomole levels of hydroperoxides using a fluorescent dichlorofluorescein assay. Anal Biochem. 1983;134(1):111-116.

   Not in our library10.1016/0003-2697(83)90270-1 (opens in a new tab)

6. 6.

   Coyle J. T., Puttfarcken P. Oxidative stress, glutamate, and neurodegenerative disorders. Science. 1993;262(5134):689-695.

   Not in our library10.1126/science.7901908 (opens in a new tab)

7. 7.

   Evans D. L., Dive C. Effects of cisplatin on the induction of apoptosis in proliferating hepatoma cells and nonproliferating immature thymocytes. Cancer Res. 1993;53(9):2133-2139.

   Not in our library

8. 8.

   Fridovich I. The biology of oxygen radicals. Science. 1978;201(4359):875-880.

   Not in our library10.1126/science.210504 (opens in a new tab)

9. 9.

   Gordon J. A., Gattone V. H. Mitochondrial alterations in cisplatin-induced acute renal failure. Am J Physiol. 1986;250(6 Pt 2):F991-F998.

   Not in our library10.1152/ajprenal.1986.250.6.f991 (opens in a new tab)

10. 10.

    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;268(27):20116-20125.

    Not in our library

11. 11.

    Matsuki N., Torii Y., Saito H. Effects of iron and deferoxamine on cisplatin-induced emesis: further evidence for the role of free radicals. Eur J Pharmacol. 1993;248(4):329-331.

    Not in our library10.1016/0926-6917(93)90008-e (opens in a new tab)

12. 12.

    McCord J. M. Oxygen-derived free radicals in postischemic tissue injury. N Engl J Med. 1985;312(3):159-163.

    Not in our library10.1056/nejm198501173120305 (opens in a new tab)

13. 13.

    Meijer C., Mulder N. H., Hospers G. A., Uges D. R., de Vries E. G. The role of glutathione in resistance to cisplatin in a human small cell lung cancer cell line. Br J Cancer. 1990;62(1):72-77.

    Not in our library10.1038/bjc.1990.232 (opens in a new tab)

14. 14.

    Meijer C., Mulder N. H., Timmer-Bosscha H., Zijlstra J. G., de Vries E. G. Role of free radicals in an adriamycin-resistant human small cell lung cancer cell line. Cancer Res. 1987;47(17):4613-4617.

    Not in our library

15. 15.

    Meister A. Mitochondrial changes associated with glutathione deficiency. Biochim Biophys Acta. 1995;1271(1):35-42.

    Not in our library10.1016/0925-4439(95)00007-q (opens in a new tab)

16. 16.

    Meister A. Selective modification of glutathione metabolism. Science. 1983;220(4596):472-477.

    Not in our library10.1126/science.6836290 (opens in a new tab)

17. 17.

    Micetich K., Zwelling L. A., Kohn K. W. Quenching of DNA:platinum(II) monoadducts as a possible mechanism of resistance to cis-diamminedichloroplatinum(II) in L1210 cells. Cancer Res. 1983;43(8):3609-3613.

    Not in our library

18. 18.

    Mickisch G. H., Roehrich K., Koessig J., Forster S., Tschada R. K., Alken P. M. Mechanisms and modulation of multidrug resistance in primary human renal cell carcinoma. J Urol. 1990;144(3):755-759.

    Not in our library10.1016/s0022-5347(17)39586-1 (opens in a new tab)

19. 19.

    Mimnaugh E. G., Dusre L., Atwell J., Myers C. E. Differential oxygen radical susceptibility of adriamycin-sensitive and -resistant MCF-7 human breast tumor cells. Cancer Res. 1989;49(1):8-15.

    Not in our library

20. 20.

    Mãrtensson J., Meister A., Mrtensson J. Glutathione deficiency decreases tissue ascorbate levels in newborn rats: ascorbate spares glutathione and protects. Proc Natl Acad Sci U S A. 1991;88(11):4656-4660.

    Not in our library10.1073/pnas.88.11.4656 (opens in a new tab)

21. 21.

    Mårtensson J., Jain A., Stole E., Frayer W., Auld P. A., Meister A. Inhibition of glutathione synthesis in the newborn rat: a model for endogenously produced oxidative stress. Proc Natl Acad Sci U S A. 1991;88(20):9360-9364.

    Not in our library10.1073/pnas.88.20.9360 (opens in a new tab)

22. 22.

    Nakashima J., Brookins J., Beckman B., Fisher J. W. Increased erythropoietin secretion in human hepatoma cells by N6-cyclohexyladenosine. Am J Physiol. 1991;261(3 Pt 1):C455-C460.

    Not in our library10.1152/ajpcell.1991.261.3.c455 (opens in a new tab)

23. 23.

    Nedwin G. E., Svedersky L. P., Bringman T. S., Palladino M. A., Goeddel D. V. Effect of interleukin 2, interferon-gamma, and mitogens on the production of tumor necrosis factors alpha and beta. J Immunol. 1985;135(4):2492-2497.

    Not in our library

24. 24.

    Sausville E. A., Stein R. W., Peisach J., Horwitz S. B. Properties and products of the degradation of DNA by bleomycin and iron(II). Biochemistry. 1978;17(14):2746-2754.

    Not in our library10.1021/bi00607a008 (opens in a new tab)

25. 25.

    Schulze-Osthoff K., Bakker A. C., Vanhaesebroeck B., Beyaert R., Jacob W. A., Fiers W. Cytotoxic activity of tumor necrosis factor is mediated by early damage of mitochondrial functions. Evidence for the involvement of mitochondrial radical generation. J Biol Chem. 1992;267(8):5317-5323.

    Not in our library

26. 26.

    Shibayama T., Tachibana M., Deguchi N., Jitsukawa S., Tazaki H. SCID mice: a suitable model for experimental studies of urologic malignancies. J Urol. 1991;146(4):1136-1137.

    Not in our library10.1016/s0022-5347(17)38025-4 (opens in a new tab)

27. 27.

    Sugihara K., Nakano S., Gemba M. Effect of cisplatin on in vitro production of lipid peroxides in rat kidney cortex. Jpn J Pharmacol. 1987;44(1):71-76.

    Not in our library10.1254/jjp.44.71 (opens in a new tab)

28. 28.

    Tietze F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem. 1969;27(3):502-522.

    Not in our library10.1016/0003-2697(69)90064-5 (opens in a new tab)

29. 29.

    Torii Y., Mutoh M., Saito H., Matsuki N. Involvement of free radicals in cisplatin-induced emesis in Suncus murinus. Eur J Pharmacol. 1993;248(2):131-135.

    Not in our library10.1016/0926-6917(93)90034-n (opens in a new tab)

30. 30.

    Uslu R., Bonavida B. Involvement of the mitochondrion respiratory chain in the synergy achieved by treatment of human ovarian carcinoma cell lines with both tumor necrosis factor-alpha and cis-diamminedichloroplatinum. Cancer. 1996;77(4):725-732.

    Not in our library

31. 31.

    Yamauchi N., Kuriyama H., Watanabe N., Neda H., Maeda M., Niitsu Y. Intracellular hydroxyl radical production induced by recombinant human tumor necrosis factor and its implication in the killing of tumor cells in vitro. Cancer Res. 1989;49(7):1671-1675.

    Not in our library

32. 32.

    Zwelling L. A., Anderson T., Kohn K. W. DNA-protein and DNA interstrand cross-linking by cis- and trans-platinum(II) diamminedichloride in L1210 mouse leukemia cells and relation to cytotoxicity. Cancer Res. 1979;39(2 Pt 1):365-369.

    Not in our library

• Anticancer activity of the iron facilitator LS081 · 2011

  Not checked

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.