LncRNA FENDRR调控miR-942-5p表达对食管癌细胞增殖、上皮间充质转化及侵袭的影响

Influences of LncRNA FENDRR on the proliferation,epithelial-mesenchymal transition and invasion of esophageal cancer cells by regulating the expression of miR-942-5p

目的:探究LncRNA FENDRR通过miR-942-5p对EC细胞增殖、上皮间充质转化(epithelial-mesenchymal transition,EMT)和侵袭的影响。方法:收集EC和食管正常组织,将EC-113细胞分为BC组(不转染)、过表达(OE)-FENDRR-NC组(转染pcDNA空质粒)、OE-FENDRR组(转染pcDNA-LncRNA FENDRR质粒)、OE-FENDRR+miR-942-5p mimics-NC组(共转染pcDNA-LncRNA FENDRR+miR-942-5p mimics-NC)和OE-FENDRR+miR-942-5p mimics组(共转染pcDNA-LncRNA FENDRR+miR-942-5p mimics)。实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)分析组织或细胞中LncRNA FENDRR、miR-942-5p表达量;细胞计数试剂盒-8(cell counting kit-8,CCK-8)法、裸鼠成瘤实验、Transwell法分析EC-113细胞体外和体内增殖和侵袭活性;双荧光素酶报告实验和RNA pull down实验分析LncRNA FENDRR和miR-942-5p的靶向关系;蛋白质印迹法(Western blot)分析EMT相关蛋白表达。结果:转染pcDNA-LncRNA FENDRR质粒可增高EC-113细胞的LncRNA FENDRR表达量,降低miR-942-5p表达,同时降低体外和体内增殖活性、侵袭数量、N-钙黏附素(N-cadherin,N-CAD)、波形蛋白(vimentin,VIM)和Snial及基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)蛋白水平,增加E-钙黏附素(E-cadherin,E-CAD)蛋白水平(P<0.05),而miR-942-5p mimics逆转了上述作用。LncRNA FENDRR可靶向并下调miR-942-5p表达。结论:LncRNA FENDRR为miR-942-5p分子海绵,可以下调miR-942-5p表达,从而抑制EC细胞增殖、EMT和侵袭能力。

Objective:To investigate the effect of lncRNA FENDRR on EC cell proliferation,epithelial-mesenchymal transition(EMT)and invasion via miR-942-5p.Methods:EC and esophageal normal tissues were collected,and EC-113 cells were separated into BC group(without transfection),overexpression(OE)-FENDRR-NC group(transfected with pcDNA empty plasmid),OE-FENDRR group(transfected with pcDNA-LncRNA FENDRR plasmid),OE-FENDRR+miR-942-5p mimics-NC group(co-transfected with pcDNA-LncRNA FENDRR+miR-942-5p mimics-NC)and OE-FENDRR+miR-942-5p mimics group(co-transfection of pcDNA-LncRNA FENDRR+miR-942-5p mimics).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was performed to analyze LncRNA FENDRR and miR-942-5p expression in tissues or cells.Cell counting kit-8(CCK-8)method,nude mouse tumorigenic experiment and Transwell method were performed to analyze the proliferation and invasion activities of EC-113 cells in vitro and in vivo.Dual-luciferase reporter assay and RNA pull down assay were performed to analyze the targeting relationship between LncRNA FENDRR and miR-942-5p.Western blot was performed to analyze EMT-related protein expression.Results:Transfection of pcDNA-LncRNA FENDRR plasmid was able to increase the expression of LncRNA FENDRR in EC-113 cells,decrease the expression of miR-942-5p,and decrease the proliferation activity and invasion number in vitro and in vivo,levels of N-cadherin(N-CAD),vimentin(VIM),snial and matrix metalloproteinase 9(MMP9)proteins,and increase the E-cadherin(E-CAD)protein level(P<0.05),while miR-942-5p mimics reversed the above effects.LncRNA FENDRR was able to target and downregulate miR-942-5p expression.Conclusion:LncRNA FENDRR is a miR-942-5p molecular sponge,which can down-regulate the expression of miR-942-5p,thereby inhibiting proliferation,EMT and invasion behavior of EC cells.