eIF4H低表达对MEL细胞增殖和红系分化的影响  

Effects of eIF4H down-expression on the proliferation and erythroid differentiation of MEL cells

作　　者：薛建有[1] 桑婷婷[1] 戚武林 曹玲玲[1] 毛群铨[1] 朱晓芳[1] 张世馥[1] 

机构地区：[1]浙江理工大学生命科学学院 蛋白质组学与分子酶学研究室,浙江杭州310018

出　　处：《基础医学与临床》2014年第2期211-215,共5页Basic and Clinical Medicine

基　　金：浙江省生物医学开放基金(SWYX0902)

摘　　要：目的探究eIF4H低表达对MEL细胞增殖和红系分化的影响。方法重组质粒载体eIF4H-shRNA-1或eIF4H-shRNA-2与pCMV-VSVG和pCMV-dR8.2共同转染HEK293T细胞,获得可使eIF4H低表达的慢病毒。慢病毒侵染MEL细胞,获得eIF4H低表达的MEL细胞稳定株,Western blot检测干扰效果。MTT检测细胞增殖;流式细胞术检测细胞周期;联苯胺染色观察红系分化情况。结果与对照组相比较,eIF4H-shRNA-2组细胞eIF4H表达明显下调,细胞增殖能力明显减弱(P<0.01),G1期细胞比例明显增加;经丁酸钠处理后eIF4H-shRNA-2组联苯胺阳性细胞明显增加(P<0.01)。结论成功构建eIF4H低表达MEL稳定株;eIF4H低表达抑制MEL细胞增殖能力;eIF4H低表达不能诱导MEL细胞红系分化,但可促进丁酸钠诱导MEL细胞红系分化。Objective To investigate the effect of eIF4H down-expression on tiation of MEL cells. Methods The eIF4H-shRNA-1 or eIF4H-shRNA-2 the proliferation and erthroid differen- plasmid, pCMV-VSVG and pCMV- dR8.2 were cotransfected into HEK293T cells to obtain Lentivirus. Lentivirus was transfected into MEL cells to es- tablish the stably eIF4H down-expressed MEL cells, Western blot was used to analyze the interference efficiency. Cells proliferation potential was measured by MTT assay; flow cytometry was used to detect the cell cycle; benzi- dine staining was applied to detect erythroid differentiation. Results Compared with the control, the expression of eIF4H was significantly down-regulated, the growth ability was significantly inhibited, the percentage of cells in G1 phase significantly increased in eIF4H-shRNA-2 cells; benzidine-positive cells showed no difference between two group cells, but after sodium butyrate treatment, benzidine-positive cells significantly increased in eIF4H-shRNA-2 cells. Conclusions We successfully established a stable eIF4H down-expressed MEL cells model; eIF4H down- expression can inhibit MEL cells proliferative capacity; MEL cells can not induce erythroid differentiation by elF4H down-expression, but promote sodium butyrate-induced erythroid differentiation of MEL cells.

关 键 词：eIF4H MEL细胞 红系分化 

分 类 号：R363[医药卫生—病理学]