circ_0044516靶向miR-1281促进食管癌细胞增殖、抑制细胞凋亡

circ_0044516 targeting miR-1281 promotes proliferation and inhibits apoptosis in esophageal cancer cells

目的:探索circ_0044516是否通过靶向miR-1281影响食管癌Eca109细胞增殖和凋亡。方法:将食管癌Eca109细胞分为si-NC组、si-circ_0044516组、miR-NC组、miR-1281组、si-circ_0044516+anti-miR-NC组和si-circ_0044516+antimiR-1281组。采用实时定量聚合酶链反应(qRT-PCR)测定食管癌组织及Eca109细胞中circ_0044516和miR-1281表达。运用CCK-8法检测Eca109细胞增殖能力,克隆形成实验测定克隆形成,Western blot测定活化的半胱氨酸蛋白酶(cleaved-caspase)3蛋白和cleaved-caspase9蛋白表达,流式细胞术测定细胞凋亡。采用荧光素酶报告实验分析circ_0044516与miR-1281的靶向结合。结果:食管癌组织中circ_0044516表达量比相匹配的癌旁组织增加2.70倍左右,miR-1281表达量较癌旁组织显著减少(P<0.05)。食管癌Eca109细胞中干扰circ_0044516表达或miR-1281过表达可使cleaved-caspase3蛋白、cleaved-caspase9蛋白表达量和凋亡率升高,细胞增殖能力降低,克隆形成数减少(P<0.05)。circ_0044516靶向调控miR-1281的表达。干扰circ_0044516表达对食管癌Eca109细胞增殖和凋亡的作用被下调miR-1281表达所逆转。结论:食管癌Eca109细胞中干扰circ_0044516表达通过靶向miR-1281抑制细胞增殖并促进其凋亡。

Objective:To explore whether circ_0044516 affects the proliferation and apoptosis of esophageal cancer Eca109cells by targeting miR-1281.Methods:Eca109 cells of esophageal cancer were divided into si-NC group,si-circ_0044516 group,miR-NC group,miR-1281 group,si-circ_0044516+anti-miR-NC group and si-circ_0044516+anti-miR-1281 group.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to determine the expression of circ_0044516 and miR-1281 in esophageal cancer tissues and Eca109 cells.CCK-8 method was used to detect the proliferation capacity of Eca109 cells,the clone formation experiment was employed to determine the clone formation,Western blot was used to measure the expressions of cleaved-cysteinyl aspartate specific proteinase(cleaved-caspase)3 protein and cleaved-caspase9 protein,and the cell apoptosis was determined by flow cytometry.The luciferase reporter experiment was used to analyze the targeted binding of circ_0044516 to miR-1281.Results:The expression of circ_0044516 in esophageal cancer tissues increased by about 2.70 times compared with the matched adjacent tissues,and the expression of miR-1281 was significantly lower than that of adjacent tissues(P<0.05).Interference with circ_0044516 expression or miR-1281 overexpression in esophageal cancer Eca109 cells increased cleaved-caspase3 protein,cleaved-caspase9 protein and apoptosis rate,and decreased cell proliferation capacity and the number of clone formation(P<0.05).circ_0044516 targeted and regulated the expression of miR-1281.The effect of interfering with circ_0044516 expression on the proliferation and apoptosis of esophageal cancer Eca109 cells was reversed by down-regulation of miR-1281 expression.Conclusion:Interference with the expression of circ_0044516 in Eca109 cells of esophageal cancer can inhibit cell proliferation and promote apoptosis by targeting miR-1281.