摘要
目的探讨Ku70乙酰化在硒诱导人肺腺癌A549细胞凋亡中的作用。方法将体外培养的人肺腺癌A549细胞分为对照组(用含体积分数10%胎牛血清的达尔伯克改良伊格尔培养基培养)和1、5、10μmol·L^(-1)硒处理组(分别用含1、5、10μmol·L^(-1)硒的达尔伯克改良伊格尔培养基培养),分别培养24、48 h,通过Western blot法检测Ku70表达,免疫沉淀法检测乙酰化Ku70和Ku70-Bax、Bax-Ku70的表达。将Ku70 siRNA转染后的人肺腺癌A549细胞分为对照组和1、5、10μmol·L^(-1)硒处理组,通过Western blot法检测凋亡相关蛋白Bax和Bcl-2的表达。结果与同一时间点对照组比较,1μmol·L^(-1)硒处理组Ku70、Bax、Bcl-2、乙酰化Ku70、Ku70-Bax和Bax-Ku70表达差异无统计学意义(P>0.05);与同一时间点对照组、1μmol·L^(-1)硒处理组比较,5、10μmol·L^(-1)硒处理组Ku70、Bcl-2、Ku70-Bax和Bax-Ku70表达均减少(P<0.01),Bax、乙酰化Ku70表达均增加(P<0.01);与同一时间点5μmol·L^(-1)硒处理组比较,10μmol·L^(-1)硒处理组Ku70、Bcl-2、Ku70-Bax和Bax-Ku70表达均减少(P<0.01),Bax、乙酰化Ku70表达均增加(P<0.01)。与本组24 h时比较,48 h时1μmol·L^(-1)硒处理组Ku70、Bax、Bcl-2、乙酰化Ku70、Ku70-Bax和Bax-Ku70表达差异无统计学意义(P>0.05);与本组24 h时比较,48 h时5、10μmol·L^(-1)硒处理组Ku70、Bcl-2、Ku70-Bax和Bax-Ku70表达减少(P<0.05),Bax、乙酰化Ku70表达增加(P<0.05)。结论硒通过促使Ku70乙酰化诱导Bax依赖性人肺腺癌A549细胞凋亡。
Objective To investigate the effect of Ku70 acetylation on selenium-induced apoptosis of human pulmonary adenocarcinoma A549 cell. Methods The A549 cells which cultured in vitro were divided into control group（ the cells were cultured in Dulbecco＇s modified Eagle＇s medium containing 10% fetal bovine serum） and 1,5,10 μmol·L^（-1）selenium group（ the cells were cultured in Dulbecco＇s modified Eagle＇s medium containing 1,5,10 μmol·L^（-1）selenium）. At the time points of 24,48 h after cultivation,the expression of Ku70 was detected by Western blotting,the acetylizad Ku70,Ku70-Bax and BaxKu70 were detected by immuno-precipitation method. The A549 cells which were transfected with Ku70 siRNA were divided into control group and 1,5,10 μmol·L^（-1）selenium group,the levels of Bax and Bcl-2 protein were detected by Western blotting. Results There was statistic difference in the level of Ku70,Bax,Bcl-2,acetylated Ku70,Ku70-Bax and Bax-Ku70 between control group and 1 μmol · L^（-1）selenium group at the same time（ P 0. 05）; Compared with control group and1 μmol·L^（-1）selenium group at the same time,5,10 μmol · L^（-1）selenium could significantly down-regulate the levels of Ku70,Bcl-2,Ku70-Bax and Bax-Ku70（ P 0. 01）,and up-regulate the levels of Bax and acetylated Ku70（ P 0. 01）; Compared with 5 μmol·L^（-1）selenium group at the same time,10 μmol·L^（-1）selenium could significantly down-regulate the levels of Ku70,Bcl-2,Ku70-Bax and Bax-Ku70（ P 0. 01）,and up-regulate the levels of Bax and acetylated Ku70（ P 0. 01）. There was no statistic difference in the levels of Ku70,Bax,Bcl-2,acetylated Ku70,Ku70-Bax and Bax-Ku70 at the time points of24 h and 48 h in 1 μmol·L^（-1）selenium group（ P 0. 05）. Compared with the time point of 24 h,the levels of Ku70,Bcl-2,Ku70-Bax and Bax-Ku70 decreased significantly（ P 0. 05）,and the levels of Bax and acetylated Ku70 increased significantly in 5,10 μmol·L^（-1）selenium groups at the time point of 48 h（ P 0. 05）. Conclusion Selenium can induce the Bax-dependent apoptosis of human pulmonary adenocarcinoma A549 cells by promoting Ku70 acetylation.
作者
刘勇
徐天娇
霍金莲
LIU Yong XU Tian-jiao HUO Jin-lian(Department of Geratology, Xianyang Hospital of Yan＇ an University, Xianyang 712000, Shaanxi Province, China Department of Pharmacology, College of Basic Medicine of Xi＇ an Medical University, Xi＇ an 710021, Shaanxi Province, China)
出处
《新乡医学院学报》
CAS
2017年第2期99-103,共5页
Journal of Xinxiang Medical University
基金
陕西省教育厅自然科学基金资助项目(编号:15JK1633)
