凋亡抑制因子（ARC）对肺癌细胞侵袭能力及对Akt蛋白磷酸化的影响

目的:探讨含有Caspase富集功能域的凋亡抑制因子（ARC）对肺癌细胞侵袭能力及对细胞中蛋白激酶B（Akt）蛋白磷酸化的影响。方法:人正常肺细胞MRC-5和肺癌细胞A549、SPC-A1、H322经细胞蛋白提取后，Western blot检测细胞中ARC表达水平。把ARC小干扰RNA组（ARC siRNA组）、siRNA对照组（siRNA-NC组）转染至人肺癌细胞中，同时设置未转染组（只加入转染试剂），培养48 h后，Western blot检测细胞中ARC、Akt、磷酸化的蛋白激酶B（p-Akt）、基质金属蛋白酶-2（MMP-2）表达水平，Transwell小室检测细胞侵袭能力。结果:人肺癌细胞A549、SPC-A1、H322中ARC表达水平明显高于正常人肺细胞MRC-5（P＜0.01）。肺癌细胞A549中ARC表达水平最高，后续选用A549细胞继续研究。ARC siRNA组细胞侵袭能力及细胞中ARC、p-Akt、MMP-2表达水平均明显低于未转染组（P＜0.01）。siRNA-NC组细胞侵袭能力及细胞中ARC、p-Akt、MMP-2表达水平与未转染组相比差异没有统计学意义（P＞0.05）。结论: ARC在肺癌细胞中表达上调，干扰ARC后肺癌细胞的侵袭受到抑制，细胞中Akt磷酸化水平降低。

The effect of ARC on invasion ability and phosphorylation of Akt protein in lung cancer cells

Objective:To investigate the effect of ARC on the invasion of lung cancer cells and the phosphorylation of Akt protein in human lung cancer cells.Methods:The expression of ARC was detected by Western blot in human normal lung cells MRC-5 and lung cancer cells A549,SPC-A1,H322 after the cell protein was extracted.siRNA-NC，ARC siRNA were transfected into human lung cancer cells，at the same time,it was set as non transfected group (transfection reagent only).After cultured 48 h,the level of ARC,Akt,p-Akt,MMP-2 were detected by Western blot,the invasion ability was detected by Transwell assay.Results:The expression of ARC in human lung cancer cells A549,SPC-A1,H322 was significantly higher than that of normal lung cells MRC-5 (P＜0.01).The expression level of ARC in lung cancer cell line A549 was the highest,and A549 cells were selected for further study.The invasion ability of ARC siRNA cells and the expression levels of ARC,p-Akt,MMP-2 in cells were significantly lower than those in transfected group (P＜0.01).There was no significant difference in ARC,p-Akt,MMP-2 expression in the cells of the siRNA-NC group compared with the non transfected group (P＞0.05).Conclusion:The invasion ability of lung cancer cells interfered with ARC expression was decreased,and MMP-2 and p-Akt were significantly decreased.