【摘要】 目的:探讨长链非编码RNA FGD5-AS1(lncRNA FGD5-AS1)对肝癌细胞放射敏感性的影响及其分子机制.方法:运用qRT-PCR检测FGD5-AS1、miR-542-3p和GTPBP4在肝癌组织和肝癌细胞系中的表达情况;采用克隆形成实验和流式细胞仪分析FGD5-AS1和GTPBP4表达变化对肝癌细胞系放射敏感性的影响.利用基因沉默、qRT-PCR和Western blot检测FGD5-AS1对miR-542-3p及其靶基因GTPBP4表达的影响,并运用荧光素酶报告实验分析lncRNA FGD5-AS1对miR-542-3p/GTPBP4表达的调控作用.结果:FGD5-AS1和GTPBP4在肝癌组织和肝癌细胞系中表达上调,miR-542-3p在肝癌组织和肝癌细胞系中表达下调.沉默FGD5-AS1可增强肝癌细胞Huh7和PLC5的放射敏感性.沉默GTPBP4可以抑制Huh7和PLC5细胞的克隆形成,促进Huh7和PLC5细胞的凋亡,增强Huh7和PLC5细胞的放射敏感性.GTPBP4是miR-542-3p的下游靶基因,miR-542-3p可以负向调控GTPBP4的表达水平;而FGD5-AS1可负向调控miR-542-3p的表达,并正向调控GTPBP4的表达水平;FGD5-AS1对HCC细胞放射敏感性的影响又依赖于miR-542-3p.结论:FGD5-AS1作为miR-542-3p的"分子海绵"竞争性上调GTPBP4的表达进而影响肝癌细胞的放射敏感性.更多还原

【Abstract】 Objective:To investigate the role and molecular mechanism of the influence of lncRNA FGD5-AS1 on radiosensitivity of hepatocellular carcinoma. Methods: qRT-PCR was used to detect the expression of FGD5-AS1, miR-542-3 p and GTPBP4 in hepatocarcinoma and hepatoma cell lines. The effects of FGD5-AS1 and GTPBP4 expression changes on the radiosensitivity of HCC cell lines were analyzed by colony formation assay and flow cytometry. The effects of FGD5-AS1 on the expression of miR-542-3 p and its target gene GTPBP4 were detected by luciferase reporter assay, gene silencing, qRT-PCR and Western blot. Analysis of the regulatory effect of lncRNA FGD5-AS1 on the expression of miR-542-3 p/GTPBP4 through the luciferase report experiment.Results: FGD5-AS1 and GTPBP4 were up-regulated in hepatocarcinoma and hepatoma cell lines, and miR-542-3 p was down-regulated in hepatocarcinoma and hepatoma cell lines. Silencing FGD5-AS1 also enhanced the radiosensitivity of Huh7 and PLC5 cells. Silencing GTPBP4 inhibited the clonal formation of Huh7 and PLC5 cells, promoted the apoptosis of Huh7 and PLC5 cells, and enhanced the radiosensitivity of Huh7 and PLC5 cells. GTPBP4 is a target gene of miR-542-3 p, and miR-542-3 p can negatively regulate the mRNA and protein expression levels of GTPBP4. However, FGD5-AS1 can negatively regulate the expression level of miR-542-3 p, and positively regulate the expression level of GTPBP4. The role of FGD5-AS1 in the radiosensitivity is dependent on miR-542-3 p. Conclusion:FGD5-AS1 can be used as a ‘molecular sponge’ of miR-542-3 p to competitively up-regulate the expression of GTPBP4 and thus affect the radiosensitivity of HCC cells.更多还原