miRNA-126靶向抑制A549细胞迁移和侵袭能力的研究  

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

作　　者：孙艳芹[1] 刘建强[2] 

机构地区：[1]广东医学院病理教研室,广东东莞523808 [2]广东医学院药学院

出　　处：《中国病原生物学杂志》2013年第10期893-895,898,共4页Journal of Pathogen Biology

基　　金：广东省自然科学基金项目(No.S2012040007835);广东医学院青年基金项目(No.XQ1116)

摘　　要：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 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.

关 键 词：非小细胞肺癌(NSCLC) EGFL7 MIR-126 

分 类 号：R392[医药卫生—免疫学]