载体介导的靶向人端粒酶反转录酶基因RNA干扰对白血病HL-60细胞增殖和凋亡的影响

Effects of vector-mediated targeting human telomerase reverse transcriptase gene RNA interference on proliferation and apoptosis of leukemia HL-60 cells

目的:探讨载体介导的靶向人端粒酶反转录酶(human telomerase reverse transcriptase,hTERT)基因RNA干扰(RNA interference,RNAi)对白血病HL-60细胞增殖和凋亡的影响。方法:采用靶向hTERT基因的RNAi重组质粒pSilencer1.0-U6/hTERT转染HL-60细胞,以空载体质粒pSilencer1.0-U6转染组、转染试剂及空白组作为对照,蛋白质印迹法检测转染后HL-60细胞hTERT蛋白的表达,MTT法检测细胞的增殖抑制率,FCM法检测HL-60细胞的凋亡情况。结果:转染pSilencer1.0-U6/hTERT质粒24、72和120h后,HL-60细胞hTERT蛋白的表达水平低于各对照组(P<0.05),细胞增殖抑制率和细胞凋亡率高于各对照组(P<0.05)。hTERT蛋白的表达水平、细胞增殖抑制率和细胞凋亡率在转染pSilencer1.0-U6/hTERT质粒24、72和120h组间无明显差异(P>0.05),在3个对照组间也无明显差异(P>0.05)。结论:载体介导的靶向hTERT基因的RNAi在体外能抑制白血病HL-60细胞hTERT蛋白的表达,抑制细胞增殖并诱导细胞凋亡。

Objective：To investigate the effects of vector-mediated targeting human telomerase reverse transcriptase（hTERT） gene RNA interference（RNAi） on the proliferation and apoptosis of leukemia HL-60 cells.Methods：The targeting hTERT gene RNAi recombination vector（pSilencer1.0-U6/hTERT） was transfected into HL-60 cells.The HL-60 cells transfected with empty vector（pSilencer1.0-U6）,RNAi-mate or HL-60 cells were used as the controls.The expression of hTERT protein was detected by Western blotting.The proliferation and apoptosis of HL-60 cells were measured by MTT assay and flow cytometry,respectively.Results：The expression levels of hTERT protein in HL-60 cells were lower and the proliferation inhibitory rate and apoptosis rate of HL-60 cells were higher than those of each control group after transfection with pSilencer1.0-U6/hTERT for 24,72,and 120 h（P0.05）.There was no differences among HL-60 cells transfected with pSilencer1.0-U6/hTERT for 24,72,and 120 h,and no differences were also observed among the three control groups（P0.05）.Conclusion：Vector-mediated targeting hTERT gene RNAi can down-regulate the expression of hTERT protein and inhibit the proliferation and induce apoptosis of HL-60 cells in vitro.