circATP8A1在肝癌中的表达及沉默其表达对肝癌细胞增殖和迁移的作用实验研究OA

Objective:To investigate the effects of silencing circular RNA(circRNA)circATP8A1 on the prolif-eration and migration of hepatocellular carcinoma cells by targeting microRNA(miR)-199a-5p.Methods:The Onco-Mine database was used to analyze the expression of circATP8A1 in liver cancer tissues and the relationship between circATP8A1 expression and survival in liver cancer patients.RT-qPCR was used to detect circATP8A1 expression in liver cancer cell lines(QGY-7701,Huh-7,SMMC-7721,HepG2,and MHCC97H).HepG2 cells were transfected with si-NC and si-circATP8A1 sequences,respectively,defining the si-NC and si-circATP8A1 groups.Colony formation assays and wound healing assays were used to assess the proliferation and migration abilities of HepG2 cells in each group.Western blot was used to detect the expression of proliferative phenotype proteins c-myc and proliferating cell nuclear antigen(PCNA),as well as migratory phenotype proteins matrix metalloproteinase-2(MMP-2)and MMP-9 in HepG2 cells from each group.Dual-luciferase reporter assays were used to verify the targeting relationship be-tween circATP8A1 and miR-199a-5p.The OncoMine database was used to analyze the correlation between cir-cATP8A1 and miR-199a-5p expression in hepatocellular carcinoma tissues.RT-qPCR was used to detect the expres-sion of miR-199a-5p in HepG2 cells from each group.Results:Expression of circATP8A1 was significantly higher in liver cancer tissues than that in normal liver tissues(P＜0.01).Hepatocellular carcinoma patients with low circATP8A1 expression had significantly longer survival than those with high circATP8A1 expression(P＜0.01).Compared with normal liver LO2 cells,circATP8A1 expression was elevated in QGY-7701,Huh-7,SMMC-7721,HepG2,and MHCC97H cells(all P＜0.01).Compared with the si-NC group,the si-circATP8A1 group showed de-creased colony formation in HepG2 cells,decreased wound healing rate,and decreased expression of c-myc,PCNA,MMP-2,and MMP-9 proteins(all P＜0.01).circATP8A1 could target and bind to miR-199a-5p.The expression of circATP8A1 was negatively correlated with miR-199a-5p in liver cancer tissues(P＜0.01).Compared with the si-NC group,the expression of miR-199a-5p in HepG2 cells in the si-circATP8A1 group was increased(P＜0.01).Conclu-sion:Silencing circATP8A1 inhibits the proliferation and migration of HepG2 liver cancer cells by targeting and up-regulating miR-199a-5p expression.