急性髓系白血病细胞中DICER1基因的表达及其功能研究  

Study on the expression and function of DICER1 in leukemic cell lines

作　　者：白颖[1] 刘彦琴[1] 王吉刚[1] 刘景华[1] 李珊珊[1] 周凡[1] 

机构地区：[1]沈阳军区总医院血液科,沈阳辽宁110056

出　　处：《现代肿瘤医学》2014年第1期22-26,共5页Journal of Modern Oncology

摘　　要：目的:检测急性髓系白血病细胞株HL-60、KG-1细胞中DICER1基因的表达水平,研究DICER1基因沉默对HL-60、KG-1细胞增殖、凋亡的影响,探寻DICER1在白血病发病机制中的作用。方法:应用Real-time PCR和Western blot检测DICER1在白血病细胞株HL-60、KG-1中mRNA和蛋白的相对表达水平。用LipofectamineTMLTX将DICER-shRNA载体转染HL-60、KG-1细胞,Real-time PCR和Western blot的方法从mRNA和蛋白水平检测DICER1的干扰效率。CCK-8法检测DICER1干扰后对白血病细胞增殖的影响,流式细胞仪检测DICER1干扰后白血病细胞凋亡率。结果:以正常HEK293细胞为对照,白血病细胞株HL-60、KG-1中DICER1 mRNA和蛋白的表达水平显著高于正常HEK293细胞(P<0.05)。DICER1-shRNA转染HL-60、KG-1细胞5天后,DICER1 mRNA和蛋白表达水平显著下降,干扰效果显著(P<0.05);CCK-8实验结果表明:与对照组细胞相比,DICER1干扰后白血病细胞增殖力显著降低(P<0.05);流式细胞分析表明:与正常细胞和对照组细胞比较,DICER1干扰后白血病细胞凋亡显著增加(P<0.01)。结论:DICER1在白血病细胞株HL-60、KG-1中高表达,具有促进白血病细胞增殖,抑制凋亡的作用。Objective：To assess the DICER1 expression levels in leukemic cell lines HL-60 and KG-1,and further investigate the effects of silencing DICER1 on cell proliferation and apoptosis in leukemia cell lines.The findings will provide a theoretical basis to reveal the role of DICER1 in the pathogenesis of leukemia.Methods：The mRNA and protein levels of DICER1 in leukemic cell lines HL-60 and KG-1 were detected by real time-PCR and Western blot.Human leukemia HL-60 and KG-1 cells were transfected with an DICER1-shRNA expressing vector by Lipofectamine^TMLTX.Real-time quantitative PCR and Western blot confirmed DICER1 knockdown in both cell lines.Cell proliferation affected by DICER1 interference was assessed using a standard CCK-8 assay.Cell apoptosis was assayed by flow cytometry.Results：DICER1 expression in leukemia cell lines HL-60 and KG-1 was significantly increased both at the mRNA and protein levels,compared with HEK cells（P〈0.05）.The level of DICER1 mRNA and protein was significantly decreased in both cell lines 5 days following transfection with DICER1-shRNA（P〈0.05）.Cell proliferation experiments revealed that DICER1 silencing inhibited cell proliferation of HL-60 and KG-1 cells（P〈0.05）.Flow cytometry analysis demonstrated that silencing of DICER1 significantly induced apoptosis compared to the controls（P〈0.01）.Conclusions：DICER1 expression level was high in leukemic cell lines HL-60 and KG-1,which may induce leukemia cell proliferation and inhibit apoptosis.

关 键 词：白血病 DICER1 HL-60细胞 KG-1细胞 增殖 凋亡 

分 类 号：R730.21[医药卫生—肿瘤] R733.71[医药卫生—临床医学]