circ-0008583结合EIF4A3蛋白促进肝癌细胞增殖并抑制其凋亡

circ-0008583 binds to EIF4A3 protein to promote the proliferation and inhibit the apoptosis of HCC cells

目的探讨circ-0008583对肝癌细胞系增殖、凋亡的影响及其作用机制。方法生物信息学分析联合临床样本和体外细胞实验验证circ-0008583在肝癌中的表达水平。RNase R和放线菌素D处理验证circ-0008583的circRNA特性。克隆形成实验检测细胞克隆形成能力,TUNEL染色检测细胞凋亡。RIP实验检测circ-0008583和EIF4A3是否结合。qRT-PCR和Western blot检测基因表达水平。结果与正常肝组织及细胞系相比,circ-0008583在肝癌组织和细胞系中高表达(均P<0.001)。circ-0008583具有环状结构,与DLG1 mRNA相比,其稳定性增加(P<0.001)。与阴性对照组相比,干扰circ-0008583表达后肝癌细胞的克隆形成能力降低(P<0.01),凋亡水平上升(P<0.001)。Circular RNA Interactome数据库和RIP实验证实circ-0008583能够与EIF4A3蛋白结合。与正常肝组织及细胞系相比,EIF4A3在肝癌组织及细胞中高表达(P<0.001)。过表达EIF4A3能够在一定程度上逆转干扰circ-0008583表达对肝癌细胞增殖的抑制作用以及凋亡的促进作用。结论circ-0008583在肝癌组织及细胞系中高表达,其与EIF4A3蛋白结合促进肝癌细胞增殖,并抑制肝癌细胞凋亡。

Objective The aim of this study was to investigate the effects and mechanism of circ-0008583 on proliferation and apoptosis of hepatoma carcinoma cell(HCC).Methods Bioinformatics analysis combined clinical samples and in vitro cell experiments were verified the expression level of circ-0008583 in HCC.RNase R and actinomycin D were used to verify the circRNA properties of circ-0008583.Cloning formation experiment was used to test cell clone formation capacity,and TUNEL staining was used to detected cell apoptosis.We used the RIP experiment to detect whether circ-0008583 could combined with EIF4 A3.qRT-PCR and Western blot were used to detect gene expression levels.Results Compared to normal liver tissues and cell lines,circ-0008583 was highly expressed in HCC tissues and cell lines(P<0.001).Circ-0008583 has a circular structure and its stability is increased compared with DLG1 mRNA(P<0.001).Compared to the negative control group,the clonal formation capacity of HCC cells after interfering circ-0008583 was reduced(P<0.01),and apoptosis level was increased(P<0.001).We use Circular RNA interactome database and RIP experiments confirm that circ-0008583 could combine with EIF4 A3 protein.Compared to normal liver tissues and cell lines,EIF4 A3 was highly expressed in HCC tissues and cell line(P<0.001).We confirmed that overexpression EIF4 A3 could reverse the inhibitory effects of circ-0008583 expression on liver cancer cell proliferation and apoptosis by rescue experiments.Conclusion Circ-0008583 is highly expressed in HCC tissues and cell lines,it combines with EIF4 A3 protein to promote HCC cell proliferation and inhibit cell apoptosis.