顺铂通过p66Shc-线粒体信号通路诱导人肾小管上皮细胞凋亡  

Cisplatin induces apoptosis of human renal tubular epithelial cells through p66^(Shc)-mitochondrial signal pathway

作　　者：聂静[1,2] 王懿春 黎祖荣[1,2] 董静 

机构地区：[1]湖南省肿瘤医院 [2]中南大学湘雅医学院附属肿瘤医院ICU,湖南长沙410013

出　　处：《中国病理生理杂志》2013年第12期2167-2171,共5页Chinese Journal of Pathophysiology

摘　　要：目的:探讨p66Shc-线粒体信号通路在顺铂诱导的人肾小管上皮细胞凋亡中的作用。方法:体外培养人肾小管上皮细胞,Western blotting法检测顺铂对p66Shc及磷酸化p66Shc(Ser36)蛋白表达的影响,然后将细胞分为对照组、顺铂组及顺铂+p66ShcS36A(第36位Ser突变为Ala的p66Shc)组,用激光共聚焦显微镜观察p66Shc对顺铂诱导的细胞活性氧簇、线粒体活性氧簇及细胞凋亡的影响,Western blotting法检测线粒体凋亡信号通路相关蛋白的表达。结果:顺铂促进p66Shc蛋白磷酸化,对p66Shc蛋白表达无影响;顺铂诱导细胞凋亡,细胞及线粒体活性氧簇产生增加,细胞色素C释放,caspase-9表达增加,p66ShcS36A可以抑制顺铂诱导的细胞氧化损伤及凋亡。结论:顺铂通过p66Shc-线粒体信号通路诱导人肾小管上皮细胞凋亡。AIM： To evaluate the roles of p66shc-mitochondrial signal pathway in apoptosis of human renal tu- bular epithelial cells induced by cisplatin. METHODS： The human renal tubular epithelial cells were cultured in vitro. The levels of p66sac and phospho-p66Shc （Ser36） protein were detected by Western blotting. The ceils were divided into con- trol group, cisplatin group and cisplatin ＋ p66ShcS36A（ p66sh~ with Ser mutating into Ala at position 36） group. The effects of p66s~ on cisplatin - induced cellular reactive oxygen species （ ROS）, mitochondrial ROS and apoptosis were measured by confocal microscopy. The expression of the proteins related to apoptosis mitochondrial signal transduction pathway was analyzed by Western blotting. RESULTS： Cisplatin induced p66she phosphorylation, but did not affect the expression of p66sh~. Cisplatin enhanced apoptosis and production of both cellular and mitochondrial ROS, release of cytochrome C and expression of caspase-9, which were inhibited by the transfection of p66ShcS36A. CONCLUSION： Cisplatin induces apop- tosis of human renaltubular epithelial cells through p66Shc-mitochondrial signal pathway.

关 键 词：顺铂 线粒体 细胞凋亡 活性氧 

分 类 号：R363[医药卫生—病理学]