摘要: |
目的:探讨微小RNA(microRNA)在叉头转录因子M1 (Fox M1)激活非小细胞肺癌(non small cell lung cancer, NSCLC)细胞上皮向间质转化(epithelial mesenehymal transition,EMT)中的作用。
方法:将Fox M1过表达质粒,Fox M1 shRNA,miR 539、miR 485 5p模拟物及其抑制物转染至人NSCLC细胞株中,应用Western印迹和real time PCR检测细胞株中Fox M1蛋白和mRNA及miR 539、miR 485 5p的表达。同时应用CCK 8和细胞迁移实验观察Fox M1、miR 539和 miR 485 5p对NSCLC细胞的增殖和侵袭功能的影响。应用荧光素酶报告基因实验检测miR 539和 miR 485 5p与EMT调控蛋白ZEB1和Snail1的关系。
结果:Fox M1过表达下调NSCLC 细胞中miR 539、miR 485 5p,转染Fox M1 shRNA后NSCLC细胞中 miR 539、miR 485 5p升高。MiR 539和miR 485 5p抑制Fox M1对NSCLC细胞增殖和侵袭的促进作用。NSCLC细胞中miR 539对EMT调控蛋白ZEB1,miR 485 5p对EMT调控蛋白Snail1的表达有抑制作用。
结论:miR 539和miR 485 5p能抑制NSCLC细胞中Fox M1 EMT通路,从而影响NSCLC细胞的增殖和侵袭,抑制NSCLC的远处转移。 |
关键词: 叉头转录因子M1 上皮向间质转化 非小细胞肺癌 微小RNA |
DOI:10.12025/j.issn.1008-6358.2016.20160990 |
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Abstract: |
Objective:To investigate the role of microRNAs in Fox M1 induced non small cell lung cancer (NSCLC) epithelial mesenchymal transition (EMT).
Methods:Over expression Fox M1 plasmid, Fox M1 shRNA, miR 539, miR 485 5p mimics and inhibitor were transfected into NSCLC cells. The expression of Fox M1 protein and mRNA, miR 539 and miR 485 5p were detected by Western blotting and real time PCR. The CCK 8 and cell migration was used to observe the effect of Fox M1, miR 539 and miR 485 5p on the proliferation and invasion of NSCLC. Luciferase reporter assay was used to explore the relationship between miRNA( miR 539 and miR 485 5p) and EMT regulatory protein (ZEB1 and Snail1).
Results:Real time RT PCR and Western blotting showed Fox M1 over expression inhibited the expression of miR 539, miR 485 5p, miR 539 and miR 485 5p in NSCLC cells were increased after transfected Fox M1 shRNA. MiR 539 and miR 485 5p suppressed enhancement of proliferation and invasion of NSCLC cells by Fox M1. miR 539 targeted and inhibited the translation of ZEB1,miR 485 5p targeted and inhibited the translation of Snail1 in NSCLC cells.
Conclusions:The mechanism of Fox M1 up regulation of EMT protein is to decrease the expression of miR 539 and miR 485 5p in NSCLC cells, thus to affect the proliferation and invasion of NSCLC cells, and to inhibit distant metastasis. |
Key words: forkhead box M1 epithelial mesenchymal transition NSCLC microRNA |