miR-539增强顺铂对非小细胞肺癌耐药株的杀伤作用研究

目的 研究miR-539对顺铂诱导的非小细胞肺癌(non-small cell lung cancer,NSCLC)耐药细胞株凋亡的影响.方法 通过荧光定量聚合酶链反应(PCR)检测NSCLC细胞系和人正常支气管上皮细胞16HBE中miR-539表达情况;通过转染miR-539建立miR-539过表达细胞系,观察miR-539过表达对顺铂抑制NSCLC细胞生长率的影响,检测细胞凋亡水平及相关基因的表达.结果 NSCLC细胞系中miR-539表达水平低于人正常支气管上皮细胞(P<0.01).阴性对照组培养48、72、96 h NCI-H460细胞增殖率均明显低于空白对照组,转染miR-539组培养不同时间NCI-H460细胞增殖率均明显低于阴性对照组和空白对照组(P<0.01).阴性对照组和转染miR-539组NCI-H460细胞凋亡率高于空白对照组(P<0.05).阴性对照组和转染miR-539组的Bcl-2、Bcl-xL基因表达量均低于空白对照组,且转染miR-539组低于阴性对照组,而Bim、Bax、P53、Myc基因表达量高于空白对照组,且转染miR-539组高于阴性对照组(P<0.05,P<0.01).结论 miR-539能够增强顺铂对NSCLC耐药细胞株的杀伤作用,可作为NSCLC基因治疗的有效靶点.

Effect of cis-Dichlorodiamineplatinum Cytotoxic Activity Enhanced by miR-539 on Non-small Cell Lung Cancer Cells

Objective To investigate the effect of miR-539 on resistance apoptosis in non-small cell lung cancer ( NSCLC) cells induced by cis dichlorodiamine platinum. Methods The miR-539 expressions in NSCLC cell line and human normal bronchial epithelial cells 16HBE were detected using fluorescent quantitation polymerase chain reaction (Q-PCR);miR-539 over-expression cell line was established by transfecting miR-539, and the effect of over-expression miR-539 on inhibiting NSCLC cell growth rate by cis dichlorodiamine platinum was observed, and the apoptosis level and expressions of relative genes were also detected. Results The miR-539 expression in NSCLC cell line was significantly decreased compared with that in human normal bronchial epithelial cells (P<0. 01). The proliferation rates of negative NCI-H460 cells 48, 72 and 96 h after cultivation were significantly lower than that in control group, and proliferation rates of NCI-H460 cells in different cultivation times in miR-539 transfection group were significantly lower than those in negative and control groups ( P<0. 01 ) . The apoptosis rates of NCI-H460 cells in negative and miR-539 transfection groups were higher than that in control group ( P<0. 05 ) . The expression levels of Bcl-2 and Bcl-xL in negative and miR-539 transfection groups were significantly lower than those in control group, and the rates in miR-539 transfection group were significantly lower than those in control group, while the Bim, Bax, P53 and Myc gene expressions were sig-nificantly higher than those in control group, and the rates in miR-539 transfection group were significantly higher than those in control group (P<0. 05, P<0. 01). Conclusion MiR-539 can enhance the lethal effect of DPP on NSCLC re-sistance cell line, and it can be used as an effective target for NSCLC gene therapy.