siRNA沉默MMP-2基因对肺癌A549细胞侵袭能力的影响

Effect of Silencing MMP-2 by siRNA on the Invasion Capability of Lung Cancer A549 Cells

目的探讨siRNA靶向抑制MMP-2基因的表达对肺癌A549细胞侵袭的抑制作用。方法化学合成一对针对MMP-2基因的干扰序列siRNA和一对阴性对照的无义序列siRNA-N,转染肺癌A549细胞,同时以未转染A549细胞作为空白对照。于转染后48 h,分别采用半定量RT-PCR技术和Western blotting法从mRNA和蛋白水平检测干扰效果;采用Boyden侵袭小室检测转染MMP-2siRNA后对A549细胞侵袭能力的影响。结果与空白对照组和转染阴性对照siRNA-N组相比,A549细胞转染MMP-2 siRNA 48 h后,MMP-2基因转录水平和蛋白水平的表达均受到抑制。转染MMP-2 siRNA组细胞穿膜数与空白对照组和转染阴性对照siRNA-N组相比明显下降。结论靶向MMP-2的siRNA不仅能特异性抑制MMP-2 mRNA和蛋白的表达,且能有效抑制A549肺癌细胞的侵袭,为肺癌的基因治疗提供了有效的靶点。

Objective To explore the inhibition effect of silencing MMP-2 by siRNA on the invasion capability of lung cancer,A549 cells in vitro.Methods siRNA targeting MMP-2 and siRNA-N as a negative control were chemically synthesized,and were transfected into A549 cells;at the same time,untransfected A549 cells was studied as a bank control.MMP-2 mRNA and protein levels were evaluated by semi-quantitative RT-PCR and Western blotting after 48 h.Boyden chamber was used to evaluate the invasion capability of A549 cells in vitro.Results Compared with two control groups,siRNA targeting MMP-2 can efficiently suppress the expressions of MMP-2 in A540 cells after 48 h.The invasion ability of A549 cells were also degraded after siRNA interference of MMP-2 gene.Conclusion siRNA targeting MMP-2 not only can degrade the expressions of MMP-2 mRNA and protein,but also can inhibit the invasion ability of A549 cells;MMP-2 may be a potentially valuable therapeutic target in the gene treatment of lung cancer.