lncRNA FAM95B1在胶质瘤中的表达及其影响LN382细胞增殖和迁移的可能机制

Expression of lncRNA FAM95B1 in glioma and the possible mechanism of its effect on the proliferation and migration of LN382 cells

目的:探讨lncRNA FAM95B1对胶质瘤细胞增殖和迁移能力的影响并探究其相关作用机制。方法:选取2018年1月至2020年8月于合肥市第三人民医院行手术治疗的38例胶质瘤患者的胶质瘤组织及癌旁组织标本,利用qPCR检测胶质瘤组织与4种细胞系中FAM95B1的表达水平,以表达最低的胶质瘤LN382细胞为研究对象,转染空载质粒(对照组)或pcDNA3.1-FAM95B1质粒(实验组)。MTT法和划痕实验检测FAM95B1对LN382细胞增殖和迁移能力的影响。生物信息学分析技术和双荧光素酶基因报告实验预测并验证FAM95B1与miR-26a-5p及PTEN之间的相互作用机制,应用qPCR和WB法检测FAM95B1对miR-26a-5p和PTEN表达的影响。结果:FAM95B1在胶质瘤组织中的表达明显低于癌旁组织(P<0.01)。FAM95B1在多种胶质瘤细胞系中的表达均明显低于正常脑胶质细胞(均P<0.01)。过表达FAM95B1可以下调LN382细胞的增殖(P<0.05)和迁移能力(P<0.01)。FAM95B1能够靶向结合miR-26a-5p(P<0.01),miR-26a-5p能够靶向结合PTEN mRNA(P<0.01)。过表达FAM95B1可下调LN382细胞中miR-26a-5p的表达(P<0.01),促进PTEN mRNA的表达(P<0.01)。结论:在胶质瘤组织和细胞系中异常低表达的FAM95B1通过发挥竞争性内源RNA的功能抑制miR-26a-5p的表达而增强PTEN蛋白表达,抑制胶质瘤细胞系LN382的增殖和迁移。

Objective:To explore the effects of long-chain non-coding RNA(lncRNA)FAM95B1 on the proliferation and migration of glioma cells and explore its related mechanisms.Methods:The glioma tissues and corresponding para-cancerous tissues of 38 glioma patients who underwent surgical treatment in The Third People's Hospital of Hefei from January 2018 to August 2020 were selected for this study.qPCR was used to detect the expression level of FAM95B1 in glioma tissues and cell lines.LN382 cells with the lowest FAM95B1 expression were used as the research object and transfected with empty plasmid(control group)or pcDNA3.1-FAM95B1 plasmid(experimental group).MTT assay and Scratch test were used to detect the effect of FAM95B1 on the proliferation and migration of LN382 cells.Bioinformatics tools were used to predict the relationship among FAM95B1,miR-26a-5p,and the phosphatase and tensin homology deleted on chromosome ten(PTEN),which was further verified by Dual-luciferase reporter gene assay.qPCR and Western blotting were used to detect the effect of FAM95B1 on the expression of miR-26a-5p and PTEN.Results:The expression of FAM95B1 in glioma tissues was significantly lower than that in para-cancerous tissues(P<0.01).The expression of FAM95B1 in various glioma cell lines was significantly lower than that in normal brain glial cells(all P<0.01).Overexpression of FAM95B1 could inhibit the proliferation(P<0.05)and migration ability(P<0.01)of LN382 cells.FAM95B1 could complementarily bind with miR-26a-5p(P<0.01),and miR-26a-5p could complementarily bind with PTEN mRNA(P<0.01).Overexpression of FAM95B1 could down-regulate the expression of miR-26a-5p(P<0.01),and promote the mRNA and protein expression of PTEN(P<0.01)in LN382 cells.Conclusion:The abnormally low expression of FAM95B1 in glioma tissues and cell lines inhibits the expression of miR-26a-5p by exerting the function of competitive endogenous RNA to enhance PTEN gene expression, and inhibits theproliferation and migration of glioma cell line LN382.