Characterization of mitochondria in cisplatin-resistant human ovarian carcinoma cells.
One of the mechanisms of cisplatin cell cytotoxicity is the mitochondria-associated induction of apoptosis. The morphological or functional change of mitochondria in cisplatin-resistant cells has already been reported. Herein we present additional data describing the mitochondrial genomic and functional changes in cisplatin- resistant cells. Cisplatin increased the level of apoptotic cells in cisplatin-sensitive human ovarian carcinoma OV 2008 and C13 cells by 3.90+/-1.01 (SD; N=3) (p<0.01)-fold and 2.03+/-0.20 (SD; N=3) (p<0.01)-fold compared to the basal apoptotic level. This indicates a lower level induction of apoptosis by 50% in cisplatin-resistant OV 2008/C13 *5.25 variant (C13) cells. In both cell types, cisplatin cytotoxicity is mostly inhibited by the caspase-9 inhibitor as well as the caspase-3 inhibitor, Ac-DEVD-CHO, suggesting that the mitochondrial downstream event was functioning well in both the C13 cells and in OV 2008 cells. Mitochondrial transmembrane potential (DeltaPsim) determined by flow cytometry using DiOC6-stained cells revealed a significant depolarization of C13 cells as compared to OV 2008 cells. Treatment of these cells with cisplatin or hydrogen peroxide induces complete mitochondrial DNA damage in OV 2008 cells, while only partial DNA-destruction is observed in C13 cells, strongly suggesting that mitochondria are resistant to cisplatin and oxidative stress response. Continuous oxygen consumption of these cells monitored by a multi-channel dissolved oxygen meter is 1.70-fold higher in OV 2008 cells than C13 cells and the oxygen consumption was decreased by 30% in C13 cells, suggesting mitochondrial respiratory malfunction in these cells. The hypothesis generated here is that mitochondrial DNA resistance to cisplatin and oxidative stress response might be one of the main characteristics concerning the lower level of apoptosis induced by cisplatin. However, the mechanism by which the mitochondrial DNA encoded molecule is involved in cisplatin resistance remains to be determined.
Hirama
Masanori
M
Department of Obstetrics/Gynecology and Department of Anatomy II, Jikei University School of Medicine, Minato-ku, Tokyo 105, Japan.
Isonishi
Seiji
S
Yasuda
Makoto
M
Ishikawa
Hiroshi
H
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D061945:Caspase Inhibitors; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D018284:Cystadenocarcinoma, Serous / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D004272:DNA, Mitochondrial / Q000235:genetics; D004347:Drug Interactions; D019008:Drug Resistance, Neoplasm; D005260:Female; D005419:Flavonoids / Q000037:antagonists & inhibitors; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009842:Oligopeptides / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000473:pathology; D010101:Oxygen Consumption / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism
D061945:Caspase Inhibitors; D004272:DNA, Mitochondrial; D005419:Flavonoids; D009842:Oligopeptides; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; D011493:Protein Kinase C; C059811:orobol; D002945:Cisplatin