Cisplatin cytotoxicity is dependent on mitochondrial respiration in Saccharomyces cerevisiae

Document Type: Original Article


1 Institute of Genetics & Hospital for Genetic Diseases, Osmania University, Hyderabad, Telangana, India-500016

2 Department of Biochemistry, Osmania University, Hyderabad, Telangana, India- 500007

3 Department of Biochemistry, Kakatiya University, Warangal, Telangana, India -506009


Objective(s): To understand the role of mitochondrial respiration in cisplatin sensitivity, we have employed wild-type and mitochondrial DNA depleted Rho0 yeast cells.
Materials and Methods: Wild type and Rho0 yeast cultured in fermentable and non-fermentable sugar containing media, were studied for their sensitivity against cisplatin by monitoring growth curves, oxygen consumption, pH changes in cytosol/mitochondrial compartments, reactive oxygen species production and respiratory control ratio.
Results: Wild-type yeast grown on glycerol exhibited heightened sensitivity to cisplatin than yeast grown on glucose. Cisplatin (100 μM), although significantly reduced the growth of wild- type cells, only slightly altered the growth rate of Rho0 cells. Cisplatin treatment decreased both pHcyt and pHmit to a similar extent without affecting the pH difference. Cisplatin dose-dependently increased the oxidative stress in wild-type, but not in respiration-deficient Rho0 strain. Cisplatin decreased the respiratory control ratio.
Conclusion: These results suggest that cisplatin toxicity is influenced by the respiratory capacity of the cells and the intracellular oxidative burden. Although cisplatin per se slightly decreased the respiration of yeast cells grown in glucose, it did not disturb the mitochondrial chemiosmotic gradient.


1. Vokes EE, Weichselbaum RR, Mick R, McEvilly JM, Haraf DJ, Panje WR. Favorable long-term survival following induction chemotherapy with cisplatin, fluorouracil, and leucovorin and concomitant chemoradiotherapy for locally advanced head and neck cancer. J Natl Cancer Inst 1992; 84:877-882.

2. Reedijk J, Lohman PH. Cisplatin: synthesis, antitumour activity and mechanism of action. Pharm Weekbl Sci 1985; 7:173-180.

3. Von Hoff DD, Schilsky R, Reichert CM, Reddick RL, Rozencweig M, Young RC, et al. Toxic effects of cis-dichlorodiammineplatinum(II) in man. Cancer Treat Rep 1979; 63:1527-1531.

4. Kartalou M, Essigmann JM. Mechanisms of resistance to cisplatin. Mutat Res 2001; 478:23-43.

5. Lindauer E, Holler E. Cellular distribution and cellular reactivity of platinum (II) complexes. Biochem Pharmacol 1996; 52:7-14.

6. Fuertes MA, Castilla J, Alonso C, Perez JM. Cisplatin biochemical mechanism of action: from cytotoxicity to induction of cell death through interconnections between apoptotic and necrotic pathways. Curr Med Chem 2003; 10:257-266.

7. Hengartner MO. The biochemistry of apoptosis. Nature 2000; 407:770-776.

8. Lemasters JJ, Qian T, Bradham CA, Brenner DA, Cascio WE, Trost LC, et al. Mitochondrial dysfunction in the pathogenesis of necrotic and apoptotic cell death. J Bioenerg Biomembr 1999; 31:305-319.

9. Olivero OA, Semino C, Kassim A, Lopez-Larraza DM, Poirier MC. Preferential binding of cisplatin to mitochondrial DNA of Chinese hamster ovary cells. Mutat Res 1995; 346:221-230.

10. Preston TJ, Abadi A, Wilson L, Singh G. Mitochondrial contributions to cancer cell physiology: potential for drug development. Adv Drug Deliv Rev 2001; 49:45-61.

11. Singh G, Maniccia-Bozzo E. Evidence for lack of mitochondrial DNA repair following cis-dichlorodiammineplatinum treatment. Cancer Chemother Pharmacol 1990; 26:97-100.

12. Szewczyk A, Wojtczak L. Mitochondria as a pharmacological target. Pharmacol Rev 2002; 54:101-127.

13. Tacka KA, Dabrowiak JC, Goodisman J, Penefsky HS, Souid AK. Effects of cisplatin on mitochondrial function in Jurkat cells. Chem Res Toxicol 2004; 17:1102-1111.

14. Harper ME, Antoniou A, Villalobos-Menuey E, Russo A, Trauger R, Vendemelio M, et al. Characterization of a novel metabolic strategy used by drug-resistant tumor cells. FASEB J 2002; 16:1550-1557.

15. Kruidering M, Van de Water B, de Heer E, Mulder GJ, Nagelkerke JF. Cisplatin-induced nephrotoxicity in porcine proximal tubular cells: mitochondrial dysfunction by inhibition of complexes I to IV of the respiratory chain. J Pharmacol Exp Ther 1997; 280:638-649.

16. Garrido N, Perez-Martos A, Faro M, Lou-Bonafonte JM, Fernandez-Silva P, Lopez-Perez MJ, et al. Cisplatin-mediated impairment of mitochondrial DNA metabolism inversely correlates with glutathione levels. Biochem J 2008; 414:93-102.

17. Turrens JF. Superoxide production by the mitochondrial respiratory chain. Biosci Rep 1997; 17:3-8.

18. Liu L, Bridges RJ, Eyer CL. Effect of cytochrome P450 1A induction on oxidative damage in rat brain. Mol Cell Biochem 2001; 223:89-94.

19. Forman HJ, Torres M. Redox signaling in macrophages. Mol Aspects Med 2001; 22:189-216.

20. Droge W. Free radicals in the physiological control of cell function. Physiol Rev 2002; 82:47-95.

21. Guidot DM, McCord JM, Wright RM, Repine JE. Absence of electron transport (Rho 0 state) restores growth of a manganese-superoxide dismutase-deficient Saccharomyces cerevisiae in hyperoxia. Evidence for electron transport as a major source of superoxide generation in vivo. J Biol Chem 1993; 268:26699-26703.

22. Barros MH, Netto LE, Kowaltowski AJ. H(2)O(2) generation in Saccharomyces cerevisiae respiratory pet mutants: effect of cytochrome c. Free Radic Biol Med 2003; 35:179-188.

23. Goldring ES, Grossman LI, Krupnick D, Cryer DR, Marmur J. The petite mutation in yeast. Loss of mitochondrial deoxyribonucleic acid during induction of petites with ethidium bromide. J Mol Biol 1970; 52:323-335.

24. Orij R, Postmus J, Ter Beek A, Brul S, Smits GJ. In vivo measurement of cytosolic and mitochondrial pH using a pH-sensitive GFP derivative in Saccharomyces cerevisiae reveals a relation between intracellular pH and growth. Microbiology 2009; 155:268-278.

25. Isonishi S, Saitou M, Yasuda M, Tanaka T. Mitochondria in platinum resistant cells. Hum Cell 2001; 14:203-210.

26. 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; 77:725-732.

27. Fendt SM, Sauer U. Transcriptional regulation of respiration in yeast metabolizing differently repressive carbon substrates. BMC Syst Biol 2010; 4:12.

28. Diwu Z, Lown JW. Photosensitization with anticancer agents. 15. Perylenequinonoid pigments as potential photodynamic therapeutic agents: formation of semiquinone radicals and reactive oxygen species on illumination. J Photochem Photobiol B 1993; 18:131-143.

29. Diwu Z, Lown JW. Photosensitization with anticancer agents. 16. The photo-oxidation of hypocrellin A. A mechanism study using 18O labelling. J Photochem Photobiol B 1993; 18:145-154.

30. Diwu Z, Zhang C, Lown JW. Photosensitization with anticancer agents. 18. Perylenequinonoid pigments as potential photodynamic therapeutic agents: preparation and photodynamic properties of amino-substituted hypocrellin derivatives. Anticancer Drug Des 1993; 8:129-143.

31. Diwu Z, Lown JW. Photosensitization with anticancer agents. 17. EPR studies of photodynamic action of hypericin: formation of semiquinone radical and activated oxygen species on illumination. Free Radic Biol Med 1993; 14:209-215.

32. Bharucha N, Kumar A. Yeast genomics and drug target identification. Comb Chem High Throughput Screen 2007; 10:618-634.

33. Blanc C, Deveraux QL, Krajewski S, Janicke RU, Porter AG, Reed JC, et al. Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells. Cancer Res 2000; 60:4386-4390.

34. Kim JS, Lee JH, Jeong WW, Choi DH, Cha HJ, Kim do H, et al. Reactive oxygen species-dependent EndoG release mediates cisplatin-induced caspase-independent apoptosis in human head and neck squamous carcinoma cells. Int J Cancer 2008; 122:672-680.

35. Park MS, De Leon M, Devarajan P. Cisplatin induces apoptosis in LLC-PK1 cells via activation of mitochondrial pathways. J Am Soc Nephrol 2002; 13:858-865.

36. Vijayalakshmi B, Sesikeran B, Udaykumar P, Kalyanasundaram S, Raghunath M. Chronic low vitamin intake potentiates cisplatin-induced intestinal epithelial cell apoptosis in WNIN rats. World J Gastroenterol 2006; 12:1078-1085.

37. Bodiga VL, Bodiga S, Surampudi S, Boindala S, Putcha U, Nagalla B, et al. Effect of vitamin supplementation on cisplatin-induced intestinal epithelial cell apoptosis in Wistar/NIN rats. Nutrition 2012; 28:572-580.

38. Kowaltowski AJ, de Souza-Pinto NC, Castilho RF, Vercesi AE. Mitochondria and reactive oxygen species. Free Radic Biol Med 2009; 47:333-343.

39. Poyton RO, Ball KA, Castello PR. Mitochondrial generation of free radicals and hypoxic signaling. Trends Endocrinol Metab 2009; 20:332-340.

40. Rosenfeld E, Beauvoit B. Role of the non-respiratory pathways in the utilization of molecular oxygen by Saccharomyces cerevisiae. Yeast 2003; 20:1115-1144.

41. Pourahmad J, Hosseini MJ, Eskandari MR, Shekarabi SM, Daraei B. Mitochondrial/lysosomal toxic cross-talk plays a key role in cisplatin nephrotoxicity. Xenobiotica 2010; 40:763-771.

42. Cummings BS, Schnellmann RG. Cisplatin-induced renal cell apoptosis: caspase 3-dependent and -independent pathways. J Pharmacol Exp Ther 2002; 302:8-17.