NK4基因联合奥沙利铂对人结肠癌细胞株HCT116凋亡和侵袭的影响

Effect of NK4 gene expression combined with oxaliplatin on apoptosis and invasion by human colorectal cancer HCT116 cells

目的探讨NK4基因联合奥沙利铂(oxaliplatin,L-OHP)在人结肠癌细胞HCT116凋亡和侵袭中的作用。方法构建NK4质粒表达载体,将NK4基因转染至HCT116细胞,采用流式细胞术分析NK4基因联合奥沙利铂对HCT116细胞凋亡的影响,Transwell小室法检测NK4基因联合奥沙利铂对该细胞侵袭的影响。采用金正均Q值法判断NK4基因联合奥沙利铂是否有协同作用;Western blot法检测不同药物作用下相关机制蛋白p-Akt的变化情况。结果在HCT116细胞中成功构建NK4质粒表达载体,NK4组m RNA表达水平高于阴性对照组(NC组)和空白组,差异有统计学意义(P<0.01)。与NC组相比,NK4基因联合奥沙利铂促细胞凋亡作用最显著(16.55%vs 46.86%,P<0.05);抑制细胞侵袭能力最强(0.9%vs 77.4%,P<0.05);金正均Q值法结果显示NK4基因联合奥沙利铂具有协同相加作用(0.85≤Q<1.15),且细胞凋亡和侵袭相关蛋白p-Akt的表达在联合用药组中显著下调,亦显示协同作用(P<0.01)。结论 NK4基因协同奥沙利铂可能通过Akt相关信号通路诱导HCT116结肠癌细胞凋亡并抑制该细胞侵袭。

Objective This study explored the effect of NK4 expression combined with oxaliplatin on apoptosis and invasion by human colorectal cancer cells（HCT116）. Methods An expression plasmid encoding the NK4 gene was built and transfected into HCT116 cells by Effectene transfection.The influence of NK4 associated with oxaliplatin on apoptosis was analyzed by flow cytometry,and the influence on invasion was analyzed by the transwell chamber method.Interaction of NK4 and oxaliplatin was evaluated using the Q value method of Zhengjun Jin.Western blotting was used to detect changes in levels of p-Akt with different drug stimulation. Results The NK4 expression plasmid was successfully built and transfected into HCT116 cells,and expression of the target gene was significantly higher than in the negative control group and blank group（P〈0.01）.Compared with the negative control group,NK4 expression combined with oxaliplatin significantly promoted apoptosis（16.55% vs 46.86%,P〈0.05）and significantly inhibited cell invasion（0.9% vs 77.4%,P〈0.05）.The Q value method indicated that the combined treatment had a synergistic effect on apoptosis and invasion（0.85≤Q〈1.15）.Expression of the apoptosis-and invasion-related protein p-Akt was significantly down-regulated in the combination group,which showed synergistic inhibition（P 〈0.01）.Conclusions NK4 expression combined with oxaliplatin can synergistically induce apoptosis and inhibit invasion by HCT116 cells via the Akt signaling pathway.