miR-200b靶向VEGF抑制肺癌细胞侵袭
miR-200b Promotes Non-small Cell Lung Cancer Cells Invasion by Targeting VEGF
罗卫民 1罗湘玉 1郭家龙 1林称意 1张军1
作者信息
- 1. 湖北医药学院附属十堰市太和医院，湖北 十堰442000
- 折叠
摘要
目的：探讨miR-200b是否通过靶向调控血管内皮生长因子( VEGF)抑制人非小细胞肺癌细胞侵袭,以揭示miR-200b的抑瘤机制。方法运用qRT-PCR检测miR-200b在非小细胞肺癌和癌旁正常肺组织中的表达；将非小细胞肺癌A549细胞分为miR-200b mimics组、miR-200b inhibitors组、scramble组、VEGF-siRNA组和control-siR-NA组,分别将 miR-200b mimics、miR-200b inhibitors、scramble、VEGF-siRNA、control-siRNA 转染于 A549细胞；采用Western blot检测A549细胞中VEGF蛋白表达,采用Transwell侵袭实验分别检测A549细胞侵袭能力。结果qRT-PCR检测结果显示,miR-200b在A549、16HBE细胞的表达分别为0.704±0.053、1.582±0.071,两者比较,差异有显著性( P<0.01)；Western blot结果显示,外源过表达miR-200b或沉默VEGF能明显下调A549细胞中VEGF蛋白的表达水平；Transwell侵袭实验显示,转染miR-200b mimics或沉默VEGF 36 h后穿过基底膜的细胞数分别为(127±6)和(136±8),与对照组(189±14)比较能明显抑制A549细胞的穿膜能力(P<0.05),而转染miR-200b inhibitors可拮抗VEGF-siRNA对A549细胞的侵袭抑制作用。结论 miR-200b通过靶向调控VEGF抑制人非小细胞肺癌细胞侵袭。
Abstract
Objective To investigate whether miR-200b promotes cell invasion of non-small cell lung cancer cells by targeting VEGF,thus to reveal molecular mechanism that miR-200b functions as a tumor suppressor in non-small cell lung cancer. Methods The expression of miR-200b in non-small cell lung cancer and normal lung tissues was detected by qRT-PCR.Non-small cell lung cancer cell line A549 were divided into miR-200b mimics group,miR-200b inhibitors group,scramble group, VEGF-siRNA group as well as control-siRNA group. A549 cells were transfected with miR-200b mimics,scramble,VEGF-siRNA,control-siRNA,respectively.The expressions of VEGF protein in A549 cells was detected by Western blotting.The invasion ability of A549 cells were evaluated by Transwell invasion assays. Results qRT-PCR showed that miR-200b was significantly down-regulated in non-small cell lung cancer tissues (0.704±0.053) than normal lung tissues (1.582±0.071).Western blot showed that the level of VEGF protein was inhibited by restored miR-200b or knock-down VEGF in A549 cells.After transfection of miR-200b mimics or knock-down VEGF for 36 h,Transwell invasion assay showed that the number of cells through the basement membrane was(127±6),(136±8),compared with the control group(189±14),it can significantly suppress the invasion of A549 cell (P<0.01).However,miR-200b inhibitors could an-tagonize the role of VEGF-siRNA in A549 cells. Conclusion miR-200b suppresses cell invasion by targeting VEGF in non-small cell lung cancer.
关键词
miR-200b/非小细胞肺癌/血管内皮生长因子/侵袭Key words
MiR-200b/non-small cell lung cancer/VEGF/invasion引用本文复制引用
基金项目
湖北省教育厅科学研究计划指导性项目(B2015477)
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据