中国药理学与毒理学杂志2016，Vol.30Issue(5)：532-538,7.DOI:10.3867/j.issn.1000-3002.2016.05.008
微RNA-129表达对食管鳞癌细胞增殖、凋亡和细胞周期的影响及其分子机制
Effect of microRNA-129 expression on proliferation,apoptosis and cell cycle of esophageal squamous cell cancer and its possible molecular mechanism
摘要
Abstract
OBJECTIVE To investigate the effect of microRNA-129(miR-129)expression on malignant phenotypes of esophageal squamous cell cancer(ESCC) cells and its possible molecular mechanisms. METHODS The constructed miR-129-overexpressed vector (pGCMV/EGFP/miR-129) and negative control vector (pGCMV/EGFP/miR-NC) were stably transfected into ESCC cell lines (Eca109 and EC9706),respectively. Quantitative real-time PCR(qRT-PCR)was performed to detect the expression of miR-129. MTT and flow cytometry(FCM)assays were performed to analyze the effects of miR-129 on proliferation, cell cycle and apoptosis of ESCC cells. Furthermore,a luciferase reporter vector with the putative B-cell lymphoma-2(Bcl-2)3′-untranslated region(pLUC/Bcl-2-3′-UTR-wt and pLUC/Bcl-2-3′-UTR-mut)was constructed to explore whether Bcl-2 was a direct target gene of miR-129 by detecting luciferase activity. Next,Western blotting was performed to detect the expression of Bcl-2, cleaved caspase 3 and total caspase 3 proteins. RESULTS Overexpression of miR-129 significantly inhibited proliferation(P<0.01),induced cell arrest in G0/G1 phase(P<0.05)and enhanced apoptosis (P<0.05)in ESCC cells. Luciferase reporter assay indicated that Bcl-2 was identified as a direct target gene of miR-129. Results of Western blotting showed that overexpression of miR-129 significantly reduced the expression of Bcl-2 protein and increased the expression of cleaved caspase 3 protein,but induced no changes in total caspase 3 protein in ESCC cells. CONCLUSION miR-129 functions as a tumor suppressor in ESCC cells by targeting Bcl-2 gene. Therefore,miR-129 will be a potential molecular target for the treatment of human ESCC.关键词
miR-129/食管鳞癌/细胞增殖/细胞凋亡/Bcl-2Key words
miR-129/esophageal squamous cell cancer/proliferation/apoptosis/Bcl-2分类
生物科学引用本文复制引用
李青峰,高燕萍,吴勉华..微RNA-129表达对食管鳞癌细胞增殖、凋亡和细胞周期的影响及其分子机制[J].中国药理学与毒理学杂志,2016,30(5):532-538,7.基金项目
国家自然科学基金 （）
