SHP-2在三氧化二砷诱导HEK293T细胞凋亡中的作用  

Role of SHP-2 in apoptosis of HEK293T cells induced by arsenic trioxide

作　　者：司云凤[1] 祝晓春[1] 孙东升[1] 齐琦[1] 王果元[1] 金成艳[1] 王丽娜[1] 徐长庆[1] 田野[1] 张力[1] 

机构地区：[1]哈尔滨医科大学病理生理学教研室,黑龙江哈尔滨150081

出　　处：《中国药理学通报》2009年第8期1086-1089,共4页Chinese Pharmacological Bulletin

基　　金：黑龙江省卫生厅资助项目(No2005-36);哈尔滨医科大学留学归国人员启动基金资助项目

摘　　要：目的探讨蛋白酪氨酸磷酸酶SHP-2在三氧化二砷(As2O3)诱导HEK293T细胞凋亡中的作用。方法将pIRES-GFP空载体、pIRES-GFP-SHP-2野生型和pIRES-GFP-SHP-2C459S突变体载体通过磷酸钙方法转染HEK293T细胞;在5μmol.L-1的As2O3孵育48 h后,用流式细胞术检测细胞凋亡率;Western blot方法检测Cyt-C、Caspase-3、p-ERK、ERK、p-JNK及JNK的表达变化。结果5μmol.L-1As2O3可诱导HEK293T细胞凋亡,SHP-2能抑制其作用;SHP-2C459S突变体组Cyt-C和Caspase-3表达均高于空载体组,SHP-2野生型组则低于空载体组;SHP-2野生型组p-ERK表达高于空载体组,SHP-2C459S突变体组则相反;p-JNK的表达,SHP-2C459S突变体组高于空载体组,而SHP-2野生型组低于空载体组。结论SHP-2可抑制As2O3诱导的HEK293T细胞凋亡,其机制可能与激活ERK和抑制JNK途径有关。Aim To investigate the role of the protein tyrosine phosphatase SHP-2 in apoptosis of HEK293T cell induced by arsenic trioxide. Methods pIERS-GFP （Empty Vector）, pIRES-SHP-2 （Wild Type） and pIRES-GFP-SHP-2C459s （Dominant Negative Mutant） were transfected into HEK293T cells;after 48 h, HEK293T cells were treated with 5 μmol.L^-1 As203 for 48 hours. Apoptosis was determined by flow cytometry. Moreover, Cyt-C, Caspasc-3, ERK, p-ERK, JNK and p-JNK were immunoblotted. Results SHP-2 potentially enhanced survival and inhibited apoptosis of HEK293 cells. The activation of Cyt-C and Caspase-3 was significantly enhanced with SHP-2 mutant ; the activation of ERK was significantly enhanced with SHP-2 wild type;phosphorylation level of JNK was significantly increased with SHP-2 mutant. Conclusion SHP-2 may inhibit apoptosis of HEK293T cells induced by arsenic trioxide and promote cell survival through the activation of ERK and JNK.

关 键 词：蛋白酪氨酸磷酸酶SHP-2 三氧化二砷 细胞凋亡 ERK JNK HEK293T细胞 细胞信号转导 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]