论文部分内容阅读
目的:探讨靶向抑制GINS2基因表达后对人早幼粒细胞白血病细胞株HL60增殖和凋亡的影响及其机制。方法:RT-PCR分别检测GINS2基因在三株白血病细胞HL60、U937、THP-1的表达。设计与合成针对GINS2基因的干扰序列,稳定转染高表达该基因的HL60细胞,荧光显微镜观察绿色荧光蛋白的表达情况;RT-PCR和Western blot分别检测各组细胞转染后mRNA和蛋白质水平;MTT法检细胞的增殖和凋亡情况;流式细胞术分析细胞的凋亡率;Western blot检测凋亡相关蛋白的表达。结果:在三株白血病细胞株中,GINS2的表达量由低到高依次是:THP-1,U937,HL60。干扰质粒转染HL60细胞后,与阴性对照组及未处理组相比,干扰组GINS2的mRNA和蛋白质水平均显著降低,且细胞的增殖能力明显受抑。同时,凋亡相关蛋白Bax表达水平显著增高而Bcl2显著降低。结论:干扰GINS2基因表达后,其mRNA和蛋白质水平均显著降低,可通过上调Bax表达,下调Bcl2表达来抑制HL60细胞增殖并促进其凋亡。
AIM: To investigate the effects of GINS2 gene targeting on the proliferation and apoptosis of human promyelocytic leukemia cell line HL60 and its mechanism. Methods: The expression of GINS2 gene in HL60, U937 and THP-1 leukemia cells was detected by RT-PCR. The interference gene of GINS2 gene was designed and synthesized. HL60 cells highly expressing the gene were stably transfected. The expression of GFP was observed by fluorescence microscopy. The mRNA and protein levels of GINS2 gene were detected by RT-PCR and Western blot respectively. The proliferation and apoptosis of cells were detected by MTT assay. The apoptosis rate of cells was analyzed by flow cytometry. The expression of apoptosis related proteins was detected by Western blot. Results: In three leukemia cell lines, the expression levels of GINS2 from low to high were THP-1, U937 and HL60. Compared with the negative control group and the untreated group, the expression of GINS2 mRNA and protein in the interference group was significantly decreased and the cell proliferation was significantly inhibited after the interference plasmid was transfected into HL60 cells. At the same time, the expression of apoptosis related protein Bax was significantly increased while Bcl2 was significantly decreased. Conclusion: The mRNA and protein levels of GINS2 gene were significantly down-regulated after interfering with GINS2 gene expression. The expression of Bax gene was down-regulated and the expression of Bcl2 protein was inhibited to inhibit the proliferation and promote the apoptosis of HL60 cells.