流式细胞术和微球技术分析应激细胞内HSP70表达  

The expression of HSP70 in cells under stress by flow cytometry and microsphere

作　　者：张慧锋[1] 张铎[1] 赵良中[1] 吕鹏[1] 张秀荣[1] 李妍[1] 

机构地区：[1]吉林医药学院,吉林吉林132013

出　　处：《吉林医药学院学报》2016年第4期241-244,共4页Journal of Jilin Medical University

基　　金：吉林教育厅项目(2015379);吉林省科技厅重大科技攻关项目(20140203012YY);国家自然科学基金项目(82102953)

摘　　要：目的建立检测细胞内HSP70的流式细胞分析技术,并应用其分析炎症、化疗药物(紫杉醇)和物理因素(热处理)对U937细胞、A549细胞和Hela细胞HSP70表达的影响。方法培养U937、A549和Hela细胞,分别用细菌脂多糖(LPS)、紫杉醇(PTX)和热应激(42℃)上述细胞;采用流式细胞术检测细胞内活性氧、细胞周期和细胞凋亡;分别采用传统的免疫印迹技术和自制液相微球结合流式细胞术来分析HSP70的表达水平。结果 LPS诱导U937细胞后,其细胞内活性氧(ROS)增加并发生凋亡,PTX处理A549细胞后,细胞发生G2/M期阻滞和凋亡,热处理导致Hela细胞发生凋亡。采用自制液相微球结合流式细胞术以及传统的免疫印迹技术,均可发现细胞内HSP70随处理因素强度或时间延长而表达增加。结论所建立的自制液相微球结合流式细胞术可用于细胞内蛋白表达量的分析。Objective To establish method to detect the expression of HSP70 in cells under stress condition.Methods U937,A549 and Hela cells were cultured in vitro.After treating by LPS,paclitaxel and heat stress,cells were collected.The content of ROS in cells,apoptosis and cell cycle were all measured by flow cytometry.The expression of HSP70 were detected by two methods,western blot or a new method using microsphere prepared in our lab.Results The ROS in U937 cells was up-regulated by LPS or PTX in a dose-dependent manner.The content of HSP70 increased depend on the time of heat stress stimulation.It was found the expression of HSP70 increased in dose or time dependently.Conclusion It is confirmed that the new method could be used to detect the expression of HSP70.

关 键 词：流式细胞术 微球 免疫印迹 热休克蛋白70 

分 类 号：R739.41[医药卫生—肿瘤]