Mitochondrial-derived ROS in edelfosine-induced apoptosis in yeasts and tumor cells  

作　　者：Hui ZHANG Consuelo GAJATE Li-ping YU Yun-xiang FANG Faustino MOLLINEDO 

机构地区：[1]Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, Changsha 410078, China [2]Department of Chemistry and Life Science, Xiangnan University, Chenzhou 423000, China [3]Cancer Research Center, Institute of Molecular and Cellular Biology of Cancer, Spanish National Research Council-University of Salamanca, Salamanca 37007, Spain [4]Research Unit, University Hospital of Salamanca, Salamanca 37007, Spain [5]Xiangnan University, Chenzhou 423000, China

出　　处：《Acta Pharmacologica Sinica》2007年第6期888-894,共7页中国药理学报（英文版）

基　　金：Project supported by grants from the Health Research Fund and European Commission(F1S-FEDER 04/0843 and 06/0813);Departmcnt of Education and Scicncc of Spain(SAF2005一04293);"Mutua Madrilena"Medical Research Foundation (FMM);'la Caixa" Foundation (BMOS一30-0);Consuelo GAlATE was supported by the "namon y Cajal" Program from the Department of Education and Science of Spain.

摘　　要：Aim： To investigate whether a similar process mediates cytotoxicity of 1-O- octadecyl-2-O-methyl-rac-glycero-3-phosphocholine （ET- 18-OCH3, edelfosine） in both yeasts and human tumor cells. Methods： A modified version of a previously described assay for the intracellular conversion of nitro blue tetrazolium to formazan by superoxide anion was used to measure the generation of reactive oxygen species （ROS）. Apoptotic yeast cells were detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay. DNA fragmentation and the generation of ROS were measured by cytofluorimetric analysis in Jurkat cells. Results： Edelfosine induced apoptosis in Saccharomyces cerevisiae, as assessed by TUNEL assay. Meanwhile, edelfosine induced a time- and concentration-dependent generation of ROS in yeasts. Rotenone, an inhibitor of the mitochondrial electron transport chain, prevented ROS generation and apoptosis in response to edelfosine in S cerevisiae. α-Tocopherol abrogated the edelfosineinduced generation of intracellular ROS and apoptosis. Edelfosine also induced an increase of ROS in human leukemic ceils that preceded apoptosis. The overexpression of Bcl-2 by gene transfer abrogated both ROS generation and apoptosis induced by edelfosine in leukemic cells. Changes in the relative mito- chondrial membrane potential were detected in both yeasts and Jurkat cells. Conclusion： These results indicate that edelfosine induces apoptosis in yeasts in addition to human tumor cells, and this apoptotic process involves mitochondria, likely through mitochondrial-derived ROS. These data also suggest that yeasts can be used as a suitable cell model in elucidating the antitumor mechanism of action of edelfosine.Aim： To investigate whether a similar process mediates cytotoxicity of 1-O- octadecyl-2-O-methyl-rac-glycero-3-phosphocholine （ET- 18-OCH3, edelfosine） in both yeasts and human tumor cells. Methods： A modified version of a previously described assay for the intracellular conversion of nitro blue tetrazolium to formazan by superoxide anion was used to measure the generation of reactive oxygen species （ROS）. Apoptotic yeast cells were detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay. DNA fragmentation and the generation of ROS were measured by cytofluorimetric analysis in Jurkat cells. Results： Edelfosine induced apoptosis in Saccharomyces cerevisiae, as assessed by TUNEL assay. Meanwhile, edelfosine induced a time- and concentration-dependent generation of ROS in yeasts. Rotenone, an inhibitor of the mitochondrial electron transport chain, prevented ROS generation and apoptosis in response to edelfosine in S cerevisiae. α-Tocopherol abrogated the edelfosineinduced generation of intracellular ROS and apoptosis. Edelfosine also induced an increase of ROS in human leukemic ceils that preceded apoptosis. The overexpression of Bcl-2 by gene transfer abrogated both ROS generation and apoptosis induced by edelfosine in leukemic cells. Changes in the relative mito- chondrial membrane potential were detected in both yeasts and Jurkat cells. Conclusion： These results indicate that edelfosine induces apoptosis in yeasts in addition to human tumor cells, and this apoptotic process involves mitochondria, likely through mitochondrial-derived ROS. These data also suggest that yeasts can be used as a suitable cell model in elucidating the antitumor mechanism of action of edelfosine.

关 键 词：EDELFOSINE alkyl-lysophospholipid analog antitumor drug apoptosis reactive oxygenspecies MITOCHONDRIA Bcl-2  yeast tumorcell 

分 类 号：R979.1[医药卫生—药品]