模拟微重力抑制K562细胞的红系分化  

Effect of simulated microgravity on erythroid differentiation of K562 cells and the mechanism

作　　者：吴彬[1] 郑磊[2] 胡秀梅[2] 高雅[1] 岳春燕[1] 洪佳琼[1] 宋洁[1] 平宝红[1] 

机构地区：[1]南方医科大学南方医院惠侨科,广东广州510515 [2]南方医科大学南方医院检验科,广东广州510515

出　　处：《南方医科大学学报》2015年第10期1422-1427,共6页Journal of Southern Medical University

基　　金：国防科工委专项课题(B3320062101);中英国际科技合作计划项目

摘　　要：目的探讨模拟微重力对K562细胞红系分化的影响及可能作用机制。方法利用第4代旋转细胞培养系统模拟微重力,联苯胺染色法检测细胞红系分化率;q RT-PCR分析GATA-1、GATA-2、Ets-1、F-actin、β-Tubulin、Vimentin基因表达水平;免疫荧光标记显示微丝、微管、中间纤维的荧光强度,Western blotting检测F-actin、β-Tubulin、Vimentin蛋白表达水平。结果联苯胺染色结果表明模拟微重力抑制K562细胞红系分化;q RT-PCR结果表明氯高铁血红素(Hemin)处理K562细胞后,调控红系分化的转录因子GATA-1表达上调,GATA-2、Ets-1表达下调;而模拟微重力处理细胞后GATA-1表达下调,GATA-2、Ets-1表达上调,细胞骨架F-actin、β-Tubulin、Vimentin表达下调;免疫荧光结果显示微丝、微管、中间纤维荧光强度减弱;免疫印迹分析结果显示Factin、β-Tubulin、Vimentin表达下调。结论模拟微重力能抑制K562细胞红系分化,其机制可能为模拟微重力破坏细胞骨架,影响GATA-1、GATA-2、Ets-1的基因表达。Objective To investigate the effect of simulated microgravity on erythroid differentiation of K562 cells and explore the possible mechanism. Methods The fourth generation rotating cell culture system was used to generate the simulated microgravity environment. Benzidine staining was used to evaluate the cell inhibition rate, and real-time quantitative PCR（q RT-PCR） was used to detect GATA-1, GATA-2, Ets-1, F-actin, β-Tubulin and vimentin m RNA expressions. The changes of cytoskeleton were observed by fluorescence microscopy, and Western blotting was employed to assay F-actin, β-tubulin and vimentin protein expression levels. Results Benzidine staining showed that simulated microgravity inhibited erythroid differentiation of K562 cells. K562 cells treated with Hemin presented with increased m RNA expression of GATA-1 and reduced GATA-2 and Ets-1 m RNA expressions. Simulated microgravity treatment of the cells resulted in down-regulated GATA-1, F-actin, β-tubulin and vimentin m RNA expressions and up-regulated m RNA expressions of GATA-2 and Ets-1, and reduced F-actin, β-tubulin and vimentin protein expressions. Exposure to simulated microgravity caused decreased fluorescence intensities of cytoskeletal filament F-actin, β-tubulin and vimentin in the cells. Conclusion Simulated microgravity inhibits erythroid differentiation of K562 cells possibly by causing cytoskeleton damages to result in downregulation of GATA-1 and up-regulation of GATA-2 and Ets-1 expressions.

关 键 词：模拟微重力 K562细胞 转录因子 红系分化 细胞骨架 

分 类 号：Q27[生物学—细胞生物学]