miRNA-126靶向抑制A549细胞迁移和侵袭能力的研究

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十勾建的miR126过表达质粒转染至NSCI．C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EGFI。7（miR126的靶标）的表达水平，采川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EGFI。7mRNA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、A549jM（）CK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〈O．05）；Transwell小室实验显示，A549／miR126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／M（）CK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EGFI。7蛋白抑制A549细胞的迁移和侵袭能力。

miR-126 may inhibit A549 cell migration and invasion by targeting EGFL7

Objective To study the impact of miR-126 on A549 cell migration and invasion. Methods A plasmid overexpressing miR 126 was constructed and transfected into A549 cells using liposomes （A549/miR 126 cells）. These cells were compared to cells transfected with an empty vector （A549/MOCK cells） and a blank control group （A549 cells）. Levels of EGFL7 （the target gene of miR-126） were detected in the three groups of cells using RT-PCR, cell mi- gration was observed using a wound healing （scratch） assay, and invasion by the three groups of cells was determined u- sing a transwell chamber assay. Results According to RT-PCR, EGFL7 mRNA was 2. 32_--4-0. 088 in A549/miR-126 cells, 1.43＋0. 026 in A549/MOCK cells, and 1.00!0. 000 in A549 cells; there were statistically significant （P~0.01） differences in the amount EGFI.7 mRNA in A549/miR-126 cells arid in the other 2 groups of cells. In the scratch assay, the average migration distance was 3.0 #m for A549/miR-126 cells, 2.65μm for A549/MOCK cells, and 0. 5 μm for A549 cells. The average inhibition rate for the three groups was 0, 11. 25%, and 83.75%, respectively. There were sta- tistically significant （P〈0.05） differences in the average migration distance and average inhibition rate for A549/miR-126 cells and the other 2 groups of cells. The transwell chamber assay indicated that 28.6＋2. 322 A549/miRq26 cells had in- vaded after 24 hr and 29.2±3. 7683 A549/miR-126 ceils had invaded after 36 hr, while 49.8±3. 7014 A549/MOCK cells had invaded after 24 hr, indicating a statistically significant decrease. Conclusion miR 126 can upregulate expression of EGFL7 mRNA and may inhibit the migration and invasion of A549 cells via EGFL7 protein, which is a product of that ex- pression.