Gene expression was assessed by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Cell growth was detected by CCK-8 and EdU assays. Flow cytometry was applied for analyzing cell apoptosis. The binding of ATP1A1-AS1 or IRF2BP2 to miR-620 was validated by RNA pulldown and luciferase reporter assays. The protein levels were examined by western blotting.
ATP1A1-AS1 was decreased in THCA cells and tissues. ATP1A1-AS1 overexpression attenuated cell growth and promoted apoptosis. MiR-620, which was upregulated in THCA, was identified as a direct target of ATP1A1-AS1. Furthermore, IRF2BP2 was discovered to be a target of miR-620, which displayed low expression in THCA cells and tissues. Importantly, IRF2BP2 knockdown reversed the influence of ATP1A1-AS1 overexpression on THCA cell proliferation and apoptosis.
LncRNA ATP1A1-AS1 inhibited cell growth and promotes apoptosis in THCA via the miR-620/IRF2BP2 axis.
方法:通过逆转录定量聚合酶链反应(RT-qPCR)评估基因表达。通过CCK-8和EdU测定检测细胞生长。流式细胞术用于分析细胞凋亡。ATP1A1-AS1或IRF2BP2与miR-620的结合通过RNA下拉和荧光素酶报告基因测定来验证。通过蛋白质印迹法检查蛋白质水平。
结果:ATP1A1-AS1在THCA细胞和组织中降低。ATP1A1-AS1过表达减弱细胞生长并促进细胞凋亡。在THCA中上调的MiR-620,被确定为ATP1A1-AS1的直接靶标。此外,IRF2BP2被发现是miR-620的靶标,其在THCA细胞和组织中显示低表达。重要的是,IRF2BP2敲低可逆转ATP1A1-AS1过表达对THCA细胞增殖和凋亡的影响。
结论:LncRNAATP1A1-AS1通过miR-620/IRF2BP2轴抑制细胞生长并促进细胞凋亡。