miR-126可靶向抑制A549细胞迁移和侵袭能力.doc

miRNA-126抑制A549细胞迁移和侵袭作用研究＊ 孙艳芹1、刘建强2 【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好的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技术检测三组细胞中miR-126及EGFL7蛋白表达水平，采用细胞划痕实验观察细胞迁移能力，Transwell小室法分析三组细胞侵袭能力的差异。结果：RT-PCR检测发现，与另两组细胞相比，A549/ miR-126组细胞中EGFL7水平较高，差异有统计学意义（P＜0.01A549/miR-126组 24h侵袭细胞数分别为(28.6±2.322)，36h侵袭细胞数为(29.2±3.7683)个，A549/mock组为(49.8±3.7014)个，差异有统计学意义（P＜0.05）。结论：miR-126可上调EGFL7的表达，抑制A549细胞的迁移和侵袭能力。 【关键词】非小细胞肺癌（NSCLC）；EGFL7；miR-126 miR-126 may inhibit A549 cell migration and invasiveness by targeted EGFL7 Sun Yanqin1, Liu Jianqiang2 [Abstract] Objective: To study the impact of miR-126 on A549 cell migration and invasiveness. Methods: Constructing miR-126 over expression plasmid and transfecting it into A549 cells by liposomes, that is A549/ miR-126 group, and set an other control group(A549 group) and an empty vector transfected group(A549/mock group). Detecting EGFL7 levels in three groups by RT-PCR, observing cell migration changes by cell scratch test, and analyzing the invasiveness ability of three groups by Transwell chamber method. Results: EGFL7 of A549/ miR-126 group significantly reduced relatively to the control groups by RT-PCR; In the cell scratch test, compared with the control group, cell migration distance of A549/ miR-126 group reduced relatively; For the Transwell chamber experiment, number of invasive A549/miR-126 cells after 24h and 36h is respectively (28.6 ± 2.322) and (29.2 ± 3.7683), while A549/mock group was (49.8-+3.7014), which was significantly reduced, the differences show statistically significant. Conclusion: miR-126 can upregulate EGFL7 expression and might inhibit the migration and invasion of A549 cells. Key words: NSCLC; EGFL7; miR-126 最新统计，在欧美发达国家和我国城市中，肺癌已位居男性恶性肿瘤发病率、死亡率的首位，女性恶性肿瘤发病率的第二位死亡率位。目前肺癌发病率呈不断上升的趋势其中占肺癌的80以上。microRNA(miRNA)是长度为18-24 nt的单链、非编码核苷酸分子，miRNA突变和异常表达与各种人体肿瘤的发生、发展关系密切，miRNA可作为癌基因或抑癌基因而起作用。作为类表皮生长因子，最初发现其在血管内皮中高表达。miR-126位于EGFL7基因3′-UTR