Troglitazone Induced Apoptosis via PPARγ Activated POX-induced ROS Formation in HT29 Cells  

作　　者：WANG Jing LV XiaoWen SHI JiePing HU XiaoSong DU YuGuo 

机构地区：[1]State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China [2]Feed Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China 3.College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China [3]College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China

出　　处：《Biomedical and Environmental Sciences》2011年第4期391-399,共9页生物医学与环境科学（英文版）

基　　金：supported by the National Basic Research Program (973) of China (No. 2009CB421605, No.2008CB418102);the National Natural Science Foundation of China (No.20890112, No. 21077127)

摘　　要：Objective In order to investigate the potential mechanisms in troglitazone-induced apoptosis in HT29 cells,the effects of PPARγ and POX-induced ROS were explored.Methods [3-（4,5）-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide （MTT） assay,Annexin V and PI staining using FACS,plasmid transfection,ROS formation detected by DCFH staining,RNA interference,RT-PCR RT-QPCR,and Western blotting analyses were employed to investigate the apoptotic effect of troglitazone and the potential role of PPARγ pathway and POX-induced ROS formation in HT29 cells.Results Troglitazone was found to inhibit the growth of HT29 cells by induction of apoptosis.During this process,mitochondria related pathways including ROS formation,POX expression and cytochrome c release increased,which were inhibited by pretreatment with GW9662,a specific antagonist of PPARγ.These results illustrated that POX upregulation and ROS formation in apoptosis induced by troglitazone was modulated in PPARγ-dependent pattern.Furthermore,the inhibition of ROS and apoptosis after POX siRNA used in troglitazone-treated HT29 cells indicated that POX be essential in the ROS formation and PPARγ-dependent apoptosis induced by troglitazone.Conclusion The findings from this study showed that troglitazone-induced apoptosis was mediated by POX-induced ROS formation,at least partly,via PPARγ activation.Objective In order to investigate the potential mechanisms in troglitazone-induced apoptosis in HT29 cells,the effects of PPARγ and POX-induced ROS were explored.Methods [3-（4,5）-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide （MTT） assay,Annexin V and PI staining using FACS,plasmid transfection,ROS formation detected by DCFH staining,RNA interference,RT-PCR RT-QPCR,and Western blotting analyses were employed to investigate the apoptotic effect of troglitazone and the potential role of PPARγ pathway and POX-induced ROS formation in HT29 cells.Results Troglitazone was found to inhibit the growth of HT29 cells by induction of apoptosis.During this process,mitochondria related pathways including ROS formation,POX expression and cytochrome c release increased,which were inhibited by pretreatment with GW9662,a specific antagonist of PPARγ.These results illustrated that POX upregulation and ROS formation in apoptosis induced by troglitazone was modulated in PPARγ-dependent pattern.Furthermore,the inhibition of ROS and apoptosis after POX siRNA used in troglitazone-treated HT29 cells indicated that POX be essential in the ROS formation and PPARγ-dependent apoptosis induced by troglitazone.Conclusion The findings from this study showed that troglitazone-induced apoptosis was mediated by POX-induced ROS formation,at least partly,via PPARγ activation.

关 键 词：TROGLITAZONE Apoptosis HT29 POX ROS formation PPARΓ Cytochrome c release 

分 类 号：Q255[生物学—细胞生物学] TQ463.5[化学工程—制药化工]