Intracellular GSH levels rather than ROS levels are tightly related to AMA-induced HeLa cell death.
Antimycin A (AMA) inhibits succinate oxidase and NADH oxidase, and also inhibits mitochondrial electron transport between cytochromes b and c. We investigated the involvement of ROS and GSH in AMA-induced HeLa cell death. AMA increased the intracellular H(2)O(2) and O(2)(*-) levels and reduced the intracellular GSH content. ROS scavengers (Tempol, Tiron, Trimetazidine and NAC) did not down-regulate the production of ROS and inhibit apoptosis in AMA-treated cells. Treatment with NAC and N-propylgallate showing the enhancement of GSH depletion in AMA-treated cells significantly intensified the levels of apoptosis. Calpain inhibitors I and II (calpain inhibitor III) and Ca(2+)-chelating agent (EGTA/AM) significantly reduced H(2)O(2) levels in AMA-treated HeLa cells. However, treatment with calpain inhibitor III intensified the levels of O(2)(*-) in AMA-treated cells. In addition, calpain inhibitor III strongly depleted GSH content with an enhancement of apoptosis in AMA-treated cells. Conclusively, the changes of ROS by AMA were not tightly correlated with apoptosis in HeLa cells. However, intracellular GSH levels are tightly related to AMA-induced cell death.
Han
Yong Hwan
YH
Department of Physiology, Medical School, Research Institute of Clinical Medicine, Center for Healthcare Technology Development, Chonbuk National University, JeonJu 561-180, Republic of Korea.
Kim
Suhn Hee
SH
Kim
Sung Zoo
SZ
Park
Woo Hyun
WH
D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt / Q000494:pharmacology; D000111:Acetylcysteine / Q000494:pharmacology; D000968:Antimycin A / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D003497:Cyclic N-Oxides / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D011435:Propyl Gallate / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013113:Spin Labels; D013481:Superoxides / Q000378:metabolism; D014292:Trimetazidine / Q000494:pharmacology
D000975:Antioxidants; D002614:Chelating Agents; D003497:Cyclic N-Oxides; D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D017382:Reactive Oxygen Species; D013113:Spin Labels; D013481:Superoxides; D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt; D004533:Egtazic Acid; D000968:Antimycin A; D011435:Propyl Gallate; C046770:EGTA acetoxymethyl ester; D006861:Hydrogen Peroxide; D053148:Caspase 3; D053181:Caspase 8; D005978:Glutathione; D014292:Trimetazidine; C001803:tempol; C058076:calpain inhibitor III; D000111:Acetylcysteine