miR-424对非小细胞肺癌细胞株A549迁移及侵袭的影响

Effect of miR-424 on migration and invasion of non-small cell lung cancer cells A549

目的探讨miR-424对非小细胞肺癌细胞株A549迁移及侵袭的影响。方法 RT-PCR法检测肺癌细胞NCI-H460、NCI-H1975、NCI-H446、A549、NCI-H1299、NCI-H157及人胚肺成纤维细胞MRC-5中miR-424表达,脂质体LipofectamineTM2000将miR-424 inhibitor和miR-424 NC转入A549细胞中,48 h后,RT-PCR法检测miR-424表达,CCK-8法检测细胞活力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,western blot检测基质金属蛋白酶2(MMP2)、MMP9、转化生长因子-β1(TGF-β1)及p-Smad3的表达。结果 miR-424在肺癌细胞NCIH460、NCI-H1975、NCI-H446、A549、NCI-H1299、NCI-H157中的[(1.78±0.13),(1.69±0.10),(1.89±0.18),(2.88±0.27),(2.52±0.20),(2.49±0.23)]表达量显著高于miR-424在人胚肺成纤维细胞MRC-5中的(0.58±0.05)表达量(P<0.01)。与miR-424 NC组比较,miR-424 inhibitor组miR-424表达量显著降低(P<0.01),细胞活力下降(P<0.01),细胞迁移及侵袭能力降低(P<0.01),MMP2,MMP9,TGF-β1及p-Smad3表达量均显著下调(P<0.01)。结论 miR-424表达量下调后能通过抑制TGF-β1/Smad3信号通路进而抑制A549细胞的迁移及侵袭。

Obejectives To explore the effect of miR-424 on migration and invasion of non-small cell lung cancer A549 cells. Methods Expression of miR-424 in NCI-H460,NCI-H1975,NCI-H446,A549,NCI-H1299,NCI-H157 lung cancer cells and embryonic lung fibroblasts MRC-5 cells was examined by RT-PCR. miR-424 inhibitor and miR-424 NC were transfected into A549 cells with liposome LipofectamineTM2000. Cell viability was measured by MTT assay. Cell migration and invasion was measured by wound scratch assay and transwell migration assay,respectively. The expression of matrix metalloproteinase 2（ MMP2）,MMP9,transforming growth factor-β（ TGF-β1）,p-Smad3 was measured by western blot. Results The expression of miR-424 in NCI-H460,NCI-H1975,NCI-H446,A549,NCI-H1299,NCI-H157 cells（ 1. 78 ± 0. 13,1. 69 ± 0. 10,1. 89 ± 0. 18,2. 88 ± 0. 27,2. 52 ± 0. 20,2. 49 ± 0. 23） was significantly higher than that inMRC-5 cell 0. 58 ± 0. 05（ P〈0. 01）. Compared with the miR-424 NC group,the cell viability was reduced（ P〈0. 01）,cell migration and invasion ability was decreased（ P〈0. 01）,and the expression of MMP2,MMP9,TGF-β1,p-Smad3 protein was down-regulated in miR-424 inhibitor group（ P〈0. 01）. Conclusions miR-424 inhibitor can inhibit the migration and invasion in lung cancer A549 cells via inhibition of TGF-β1/Smad3 signal pathway.