lncRNA TOPORS-AS1靶向miR-718对胃癌细胞增殖、迁移和凋亡调控作用研究

The regulatory effects of lncRNA TOPORS-AS1 targeting miR-718 on the proliferation,migration and apoptosis of gastric cancer

目的探讨lncRNA TOPORS-AS1靶向miR-718对胃癌细胞增殖、迁移和凋亡的调控作用。方法RT-qPCR分析TOPORS-AS1和miR-718在胃癌组织、细胞系中的表达。将pcDNA、pcDNA-TOPORS-AS1、anti-miR-NC、anti-miR-718、pcDNA-TOPORS-AS1+miR-NC、pcDNA-TOPORS-AS1+miR-718分别转染HGC-27细胞,通过CCK-8和克隆形成实验评估细胞增殖;流式细胞术检测细胞凋亡;Transwell法分析细胞迁移。TOPORS-AS1对miR-718的靶向调控作用通过RT-qPCR和双荧光素酶报告实验验证。结果TOPORS-AS1在胃癌组织、细胞系中表达显著升高(P<0.05),miR-718的表达显著降低(P<0.05)。过表达TOPORS-AS1后HGC-27细胞克隆形成数、迁移数、miR-718表达显著降低(P<0.05),抑制率、凋亡率显著升高(P<0.05)。抑制miR-718表达后HGC-27细胞克隆形成数、迁移数显著降低(P<0.05),抑制率、凋亡率显著升高(P<0.05)。miR-718是TOPORS-AS1的直接靶点。过表达miR-718显著减弱TOPORS-AS1过表达对HGC-27细胞增殖、迁移和凋亡的影响(P<0.05)。结论TOPORS-AS1负调控miR-718抑制胃癌细胞增殖、迁移并促进凋亡。

Objective To investigate the regulatory effects of lncRNA TOPORS-AS1 targeting miR-718 on the proliferation,migration and apoptosis of gastric cancer.Methods The expression levels of TOPORS-AS1 and miR-718 in gastric cancer tissues and cell lines were detected by RT-qPCR.And pcDNA,pcDNA-TOPORS-AS1,anti-miR-NC,anti-miR-718,pcDNA-TOPORS-AS1+miR-NC,pcDNA-TOPORS-AS1+miR-718 were transfected into HGC-27 cells respectively.CCK-8 and clone formation assays were performed to evaluate the cell proliferation;flow cytometry detected cell apoptosis;Transwell method was used to test cell migration.The targeted regulation effects of TOPORS-AS1 on miR-718 were verified by RT-qPCR and dual luciferase reporter experiment.Results The expression levels of TOPORS-AS1 in gastric cancer tissues and cell lines were significantly decreased(P<0.05),however,the expression levels of miR-718 were significantly decreased(P<0.05).After overexpression of TOPORS-AS1,the clone formation number,migration number and the expression levels of miR-718 in HGC-27 cells were significantly increased(P<0.05),however,the inhibition rate and apoptosis rate were significantly increased(P<0.05).After inhibiting the expression of miR-718,the clone formation number and migration number in HGC-27 cells were significantly decreased(P<0.05),however,the inhibition rate and apoptosis rate were significantly increased(P<0.05).Moreover miR-718 was the direct target of TOPORS-AS1.The overexpression of miR-718 significantly decreased the effects of TOPORS-AS1 overexpression on the proliferation,migration and apoptosis of HGC-27 cells(P<0.05).Conclusion TOPORS-AS1 can inhibit the cell proliferation,migration and promote apoptosis of gastric cancer by negatively regulating miR-718.