Enhancement of arsenic trioxide-induced apoptosis in HeLa cells by diethyldithiocarbamate or buthionine sulfoximine.
Arsenic trioxide (ATO) affects many biological functions such as cell proliferation, apoptosis, differentiation and angiogenesis in various cells. We investigated the in vitro effects of ATO as a reactive oxygen species (ROS) generator or a glutathione (GSH) depletor on apoptosis in HeLa cells. ATO decreased the viability of HeLa cells in a dose-dependent manner with an IC50 of approximately 5-6 microM. ATO triggered apoptosis, which is accompanied by the loss of mitochondrial transmembrane potential (DeltaPsim). Intracellular general ROS levels in HeLa cells were increased or decreased depending on the concentration of ATO. Particularly, the levels of O2.- were increased by ATO. In addition, we detected a decreased GSH content in ATO-treated cells. The GSH-depleted cells mainly showed propidium iodine-positive staining, indicating that the majority of the cells were dead. Diethyldithiocarbamate (DDC; an inhibitor of Cu,Zn-SOD) induced apoptosis and the loss of mitochondrial transmembrane potential (DeltaPsim) in HeLa control cells. DDC intensified apoptosis, the loss of mitochondrial transmembrane potential, increased levels of O2.- and GSH depletion in ATO-treated cells. L-buthionine sulfoximine (BSO; an inhibitor of GSH synthesis) did not induce apoptosis in HeLa control cells, but increased levels of apoptosis, O2.- and GSH depletion in ATO-treated cells. In conclusion, the changes in intracellular GSH levels rather than ROS levels are tightly related to the enhancement of ATO-induced apoptosis in HeLa cells by DDC or BSO.
Department of Physiology, Medical School, Institute for Medical Sciences, Chonbuk National University, JeonJu 561-180, Korea.
Yong Hwan
YH
Han
Sung Zoo
SZ
Kim
Suhn Hee
SH
Kim
Woo Hyun
WH
Park
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology*; D019328:Buthionine Sulfoximine / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004050:Ditiocarb / Q000494:pharmacology*; D005978:Glutathione / Q000096:biosynthesis; D006367:HeLa Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010087:Oxides / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
D000970:Antineoplastic Agents; D001152:Arsenicals; D010087:Oxides; D017382:Reactive Oxygen Species; D013481:Superoxides; D019328:Buthionine Sulfoximine; D004050:Ditiocarb; D005978:Glutathione; D000077237:Arsenic Trioxide