人表皮生长因子受体-2 siRNA联合卡铂对肺腺癌A549细胞株的抑制作用

THE INHIBITION OF A549 CELLS BY SIRNA TARGETING C-ERBB-2 GENE COMBINED WITH CARBOPLATIN

目的:应用人表皮生长因子受体-2(C-erbB-2)基因的小干扰RNA(small interfering RNA,siRNA)联合卡铂作用于肺腺癌A549细胞株,探讨siRNA敲除C-erbB-2基因联合卡铂对A549细胞的抑制作用,为开拓肺腺癌患者治疗的新途径提供实验依据。方法:应用CCK-8比色法检测不同浓度的卡铂在不同时间点对A549细胞的抑制率。设计并合成3对C-erbB-2siRNA(2621组、1724组、1491组),分别转染肺腺癌A549细胞,应用实时荧光定量RT-PCR检测该基因的mRNA水平,筛选出最佳siRNA。再应用CCK-8比色法检测卡铂与C-erbB-2 siRNA联合应用时A549细胞的抑制率。结果:卡铂对A549细胞的生长有显著抑制作用。随着浓度的升高(r=0.415,P<0.001)、作用时间的延长(r=0.689,P<0.001),细胞的抑制率升高。48h的IC50为25.99mg/L。3对siRNA中2621组和1724组干扰效果最佳(F=11.854,P<0.05)。CCK-8比色法检测显示,A549细胞的抑制率分别为:siRNA干扰组(28.68±1.98)%、卡铂组(45.63±2.14)%、siRNA+卡铂组(59.62±1.35)%,其中siRNA+卡铂组抑制率最高,与其它各组比较差异有统计学意义(P<0.01)。结论:C-erbB-2 siRNA与卡铂具有协同作用,能抑制A549细胞增殖。

Objective：To investigate the inhibition of human lung adenocarcinoma cell line A549 by siRNA targeting C-erbB-2 gene combined with carboplatin（CBP） and to provide experimental basis for the clinical treatment of lung cancer patients. Methods： CCK-8 was used to detect the inhibitory rate of A549 cells by CBP at different concentration and at diffferent time point. 3 pairs of C-erbB-2 siRNA（group 2621,group 1724 and group 1491） were designed and composed. The siRNA was transfected into A549 cells, and the interference efficincy was tested by real-time fluorescent quantitation RT-PCR to select the best C-erbB-2 siRNA. Anti-proliferative effect on A549 cells by CBP and C-erbB-2 siRNA was detected by CCK-8 assay. Results：The inhibitory rate of cells by CBP increased along with the increase of concentration（r=0.415, P〈0.001）and time（r=0.689,P〈0.001）. A- mong the 3 pairs of siRNA,group 2621 and group 1724 were more effective（F=11.854,P〈0.05）.The anti-proliferative rates of the cells in each group were： （28.68±1.98）% in interference group, （45.63±2.14）% in CBP group, and （59.62±1.35）% in interferenee＋CBP group,respectively. The anti-proliferative rate of the last group was the high- est;there was significant statistic difference （P〈0.01）. Conclusion： SiRNA targeting C-erbB-2 gene combined with CBP can improve the anti-proliferation of A549 cells.