miR-539对非小细胞肺癌细胞增殖凋亡的影响及机制

Effect of miR-539 on apoptosis related genes of non-small cell lung cancer cell line H460 and its mechanism

目的探究分析非小细胞肺癌细胞H460中miR-539表达差异及凋亡相关基因的影响,阐明其作用机制。方法采用实时荧光定量PCR(qPCR)检测非小细胞癌细胞系H460和健康人体的支气管上皮细胞(16HBE)miR-539表达。实验分为空白对照组、阴性对照组及转染组(miR-539);MTT法检测各组细胞的增殖作用,流式细胞术检测各组细胞凋亡情况,qPCR检测细胞凋亡相关基因表达。结果健康人体支气管上皮细胞的miR-539表达水平为1.01±0.09,高于NCI-H460的0.21±0.03,差异具有统计学意义(P<0.05)。48、72、96 h时,空白组和阴性对照组细胞增殖能力高于转染组;空白组细胞增殖能力高于阴性对照组(P<0.05)。转染组、阴性对照组和空白组细胞凋亡率分别为(88.4±5.7)%、(64.5±8.2)%和(58.1±5.2)%,差异有统计学意义(P<0.05)。转染组、阴性对照组和空白组细胞凋亡率分别为(88.4±5.7)%、(64.5±8.2)%和(58.1±5.2)%,差异有统计学意义(P<0.05)。转染组Bcl-xL、Bcl-2基因表达水平低于阴性对照组和空白组(P<0.05),Bax、Bim、Myc、P53基因表达水平高于阴性对照组和空白组(P<0.05)。结论miR-539在非小细胞肺癌细胞中低表达,上调miR-539表达可增强顺铂对肺癌细胞的增殖抑制作用以及凋亡水平。

Objective To explore and analyze the expression difference of miR-539 and the effect of apoptosis-related genes in non-small cell lung cancer cell H460,and to clarify its mechanism.Methods Real-time fluorescence quantitative PCR(qPCR)was used to detect the expression of miR-539 in non-small cell carcinoma cell line H460 and bronchial epithelial cells(16HBE)of healthy human.The experiment was divided into blank control group,negative control group and transfection group(miR-539).MTT assay was used to detect cell proliferation,flow cytometry was used to detect cell apoptosis,and qPCR was used to detect apoptosis-related gene expression.Results The expression level of miR-539 in healthy human bronchial epithelial cells was 1.01±0.09,higher than that of NCI-H460(0.21±0.03),the difference was statistically significant(P<0.05).At 48,72 and 96 h,the proliferation ability of blank group and negative control group was higher than that of transfection group.The proliferation ability of blank group was higher than that of negative control group(P<0.05).The apoptosis rates of transfection group,negative control group and blank group were(88.4±5.7)%,(64.5±8.2)%and(58.1±5.2)%,respectively,with statistical significance(P<0.05).The apoptosis rates of transfection group,negative control group and blank group were(88.4±5.7)%,(64.5±8.2)%and(58.1±5.2)%,respectively,with statistical significance(P<0.05).The expression levels of Bcl-xL and Bcl-2 genes in the transfected group were lower than those in the negative control group and the blank group(P<0.05),and the expression levels of Bax,Bim,Myc and P53 genes were higher than those in the negative control group and the blank group(P<0.05).Conclusion miR-539 is underexpressed in non-small cell lung cancer cells,and up-regulation of miR-539 expression can enhance the inhibitory effect of cisplatin on proliferation and apoptosis of lung cancer cells.