南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (10): 1422-.

• • 上一篇    下一篇

模拟微重力抑制K562细胞的红系分化

吴彬，郑磊，胡秀梅，高雅，岳春燕，洪佳琼，宋洁，平宝红   

  • 出版日期:2015-10-20 发布日期:2015-10-20

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

  • Online:2015-10-20 Published:2015-10-20

HTML

PDF

1008

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；qRT-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；qRT-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
F-actin、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。

Abstract: 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
(qRT-PCR) was used to detect GATA-1, GATA-2, Ets-1, F-actin, β-Tubulin and vimentin mRNA 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 mRNA expression of GATA-1 and
reduced GATA-2 and Ets-1 mRNA expressions. Simulated microgravity treatment of the cells resulted in down-regulated
GATA-1, F-actin, β-tubulin and vimentin mRNA expressions and up-regulated mRNA 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
down-regulation of GATA-1 and up-regulation of GATA-2 and Ets-1 expressions.