RNA干扰Apg-2基因表达对BaF3-MIGR1和BaF3-p210细胞凋亡的影响

Effect of interference of Apg-2 gene expression by RNA on apoptosis of BaF3-MIGR1 and BaF3-p210 cells

目的探讨RNA干扰Apg-2基因表达对BaF3-MIGR1和BaF3-p210(Bp210)细胞凋亡的影响。方法取BaF3-MIGR1和Bp210、稳定抑制Apg-2表达的BaF3-MIGR1-Hspa42和Bp210-Hspa42细胞及阴性对照Bp210-HspaHK和BaF3-MIGR1-HspaHK细胞,采用瑞特染色法观察各组细胞的形态学变化;流式细胞术检测各组细胞的凋亡情况;Western blot法检测各组细胞中凋亡相关蛋白Bax和Bcl-2的表达水平。结果干扰Apg-2基因表达后,细胞出现肿胀、胞浆空泡、核染色质聚集、核碎裂,部分细胞可见凋亡小体,处于有丝分裂中期细胞明显减少。与BaF3-MIGR1和Bp210细胞相比,BaF3-MIGR1-Hspa42和Bp210-Hspa42细胞的凋亡率显著增加(P<0.05);Bcl-2蛋白的表达水平下调,Bax蛋白的表达水平无明显变化。结论干扰Apg-2基因表达可促进BaF3-MIGR1和Bp210细胞凋亡增加,其可能是通过降低抗凋亡蛋白Bcl-2的表达水平来实现的。

Objective To investigate the effect of interference of Apg-2 gene expression by RNA on the apoptosis of BaF3-MIGR1 and BaF3-p210（Bp210） cells.Methods BaF3-MIGR1 and Bp210 cells,BaF3-MIGR1-Hspa42 and Bp210-Hspa42 cells in which Apg-2 expression was inhibited stably,as well as Bp210-HspaHK and BaF3-MIGR1HspaHK cells as negative control were observed for morphologies by Wright staining,for apoptosis by flow cytometry,and for expression of apoptosis-related Bax and Bcl-2 proteins by Western blot.Results A series of morphological changes including swelling,cytoplasmic vacuoles,chromatin aggregation,nuclear fragmentation were observed in the cells after interference of Apg-2 gene expression,and apoptotic body formation in partial cells.However,the cells at metaphase decreased significantly in quantity（P ＆lt; 0.05）.As compared with those of BaF3-MIGR1 and Bp210 cells,the apoptotic rates of BaF3-MiGR1-Hspa42 and Bp210-Hspa42 cells increased significantly（P ＆lt; 0.05）,while the Bcl-2 expression level decreased,and Bax expression level showed no significant change.Conclusion Interference of Apg-2 gene expression promoted the apoptosis of BaF3-MIGR1 and Bp210 cells through a possible mechanism of down-regulating the expression of anti-apoptotic protein Bcl-2.