摘要
目的 探讨长链非编码RNA溶质载体家族16成员1反义RNA 1(lncRNA SLC16A1-AS1)调节微小RNA-532-3p(miR-532-3p)/肌动蛋白结合蛋白2(TAGLN2)信号通路对脑胶质瘤细胞增殖、迁移和侵袭的影响及其机制.方法 实时荧光定量PCR检测胶质瘤细胞中lncRNA SLC16A1-AS1表达水平,筛选最佳干预细胞系;通过荧光原位杂交实验、下拉实验、双荧光素酶报告基因实验验证lncRNA SLC16A1-AS1、TAGLN2与miR-532-3p的靶向调控关系.①将U251细胞分为小干扰RNA阴性对照(si-NC)组、si-SLC16A1-AS1组、si-SLC16A1-AS1+NC inhibitor组、si-SLC16A1-AS1+ miR-532-3p inhibitor组、miR-NC组、miR-532-3p mimics组、miR-532-3p mimics+pcDNA组、miR-532-3p mimics+TAGLN2组:检测U251细胞的增殖、迁移和侵袭变化;Western blot检测TAGLN2、E-钙黏附蛋白、波形蛋白表达水平.②小鼠体内肿瘤形成实验:验证lncRNA SLC16A1-AS1对胶质瘤生长的影响.结果 lncRNA SLC16A1-AS1在胶质瘤细胞中表达水平升高(P＜0.05);U251细胞FISH实验、下拉实验、双荧光素酶基因实验证实lncRNA SLC16A1-AS1,TAGLN2与miR-532-3p存在靶向调控关系;抑制lncRNA SLC16A1-AS1表达或过表达miR-532-3p可显著抑制U251细胞增殖、迁移、侵袭及上皮间质转化(P＜0.05);抑制miR-532-3p表达或过表达TAGLN2可逆转抑制lncRNA SLC16A1-AS1表达或过表达miR-532-3p对U251细胞增殖、迁移、侵袭及上皮间质转化的抑制作用(P＜0.05).小鼠体内实验显示,抑制lncRNA SLC16A1-AS1表达可显著抑制移植瘤的生长(P＜0.05).结论 lncRNA SLC16A1-AS1在胶质瘤细胞中表达水平上调,抑制lncRNA SLC16A1-AS1表达可通过调节miR-532-3p/TAGLN2信号通路,进而抑制胶质瘤的恶性进展.
Abstract
Aim To investigate the impacts and mechanism of long non coding RNA solute carrier family 7,member 11 antisense RNA(lncRNA SLC16A1-AS1)on the proliferation,migration and invasion of glioma cells by regulating the microRNA-532-3p(miR-532-3p)/transgelin 2(TAGLN2)axis.Methods The expression of lncRNA SLC16A1-AS1 in glioma cells was detected by qRT-PCR to screen the best intervention cell line.The targeted regulation relationship between lncRNA SLC16A1-AS1,TAGLN2 and miR-532-3p was verified by fluorescence in situ hybridization(FISH)experiment,pull-down experiment and double luciferase reporter gene experiment.U251 cells were selected for intervention and grouped into si-NC group,si-SLC16A1-AS1 group,si-SLC16A1-AS1+NC inhibitor group,si-SLC16A1-AS1+miR-532-3p inhibitor group,miR-NC group,miR-532-3p mimics group,miR-532-3p mimics+pcDNA group,and miR-532-3p mimics+TAGLN2 group,the proliferation,migration and invasion of U251 cells were detected.Western blot was applied to detect the expression of TAGLN2,E-cadherin and vimentin;and the effect of lncRNA SLC16A1-AS1 on the growth of glioma was verified by the tumorigenesis experiment in mice.Results The expression level of lncRNA SLC16A1-AS1 in glioma cells increased(P＜0.05),U251 cells were selected for the experiment.FISH test,pull-down test and double luciferase reporter gene test confirmed that lncRNA SLC16A1-AS1,TAGLN2 and miR-532-3p had a targeted regulatory relationship.Inhibition of lncRNA SLC16A1-AS1 expression or overexpression of miR-532-3p obviously inhibited proliferation,migration,invasion and EMT of U251 cells(P＜0.05).Inhibition of miR-532-3p expression or overexpression of TAGLN2 could reverse the inhibition of lncRNA SLC16A1-AS1 expression or overexpression of miR-532-3p on proliferation,migration,invasion and epithelial-mesenchymal transition of U251 cells(P＜0.05).In vivo experiments showed that inhibiting the expression of lncRNA SLC16A1-AS1 could obviously inhibit the growth of transplanted tumor in mice(P＜0.05).Conclusion lncRNA SLC16A1-AS1 is up-regulated in glioma cells.Inhibiting the expression of lncRNA SLC16A1-AS1 can inhibit the malignant progression of glioma by regulating the miR-532-3p/TAGLN2 signal axis.
基金项目
2021年度河北省医学科学研究课题计划(20210971)
NETL
NSTL
万方数据