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目的研究microRNA-126(miR-126)对非小细胞肺癌(NSCLC)细胞迁移和侵袭能力的影响。方法利用脂质体介导法将构建的miR-126过表达质粒转染至NSCLC细胞株A549细胞(A549/miR-126组),并设空质粒转染组(A549/MOCK组)和空白对照组(A549组)。利用RT-PCR技术检测3组细胞中EGFL7(miR-126的靶标)的表达水平,采用细胞划痕实验观察细胞迁移能力差异,采用Transwell小室法分析3组细胞侵袭能力的差异。结果 RT-PCR检测A549/miR-126组、A549/MOCK组和A549组细胞中EGFL7mRNA分别为2.32±0.088、1.43±0.026和1.00±0.000,差异有统计学意义(P<0.01);细胞划痕实验显示A549/miR-126组、A549/MOCK组和A549组细胞平均迁移距离分别为3.0μm、2.65μm和0.5μm,平均抑制率分别为0、11.25%和83.75%,差异有统计学意义(P<0.05);Transwell小室实验显示,A549/miR-126组24hr侵袭细胞数为(28.6±2.322)个,36h侵袭细胞数为(29.2±3.7683)个,A549/MOCK组24h为(49.8±3.7014)个,差异有统计学意义(P<0.05)。结论 miR-126可上调EGFL7mRNA的表达,并可能通过表达产物EGFL7蛋白抑制A549细胞的迁移和侵袭能力。
Objective To investigate the effect of microRNA-126 (miR-126) on the migration and invasion of non-small cell lung cancer (NSCLC) cells. Methods The constructed miR-126 overexpression plasmid was transfected into A549 / miR-126 cells (A549 / miR-126 group) by liposome-mediated method and transfected into empty plasmid transfected A549 / MOCK group and blank control Group (A549 group). The expression of EGFL7 (the target of miR-126) in three groups of cells was detected by RT-PCR. The difference of cell migration was observed by cell scratch assay. The invasion ability of the three groups was analyzed by Transwell chamber assay. Results The mRNA of EGFL7 in A549 / miR-126 group, A549 / MOCK group and A549 group was 2.32 ± 0.088, 1.43 ± 0.026 and 1.00 ± 0.000 respectively by RT-PCR, the difference was statistically significant (P <0.01) The results showed that the average migration distances of A549 / miR-126, A549 / MOCK and A549 groups were 3.0μm, 2.65μm and 0.5μm, respectively. The average inhibition rates were 0, 11.25% and 83.75%, respectively P <0.05). Transwell chamber assay showed that the number of invasive cells in 24hr was (28.6 ± 2.322) in A549 / miR-126 group and (29.2 ± 3.7683) in 36h and 49.8 ± 3.7014 in A549 / MOCK group ), The difference was statistically significant (P <0.05). Conclusion miR-126 can up-regulate the expression of EGFL7mRNA, and may inhibit the migration and invasion ability of A549 cells through the expression of EGFL7 protein.