miR-139通过沉默B细胞转位基因3(BTG3)抑制肝细胞癌细胞的增殖

microRNA-139 suppresses proliferation of hepatocellular carcinoma cells by silencing of B cell translocation gene 3

目的初步阐明miR-139在肝细胞癌(HCC)发展过程中的作用及其介导的信号通路。方法通过CCK-8实验验证miR-139对HCC增殖的影响;运用Target Scan、MiRanda、Clip-seq和miRDB四组数据库对miR-139下游可能存在的通路进行预测,结合文献回顾,寻找出miR-139在HCC中可能存在的靶点。Western blot法验证靶点受miR-139调节的情况,双荧光素酶报告基因分析验证miR-139与靶点的相关性。结果通过CCK-8实验证明,miR-139具有抑制肝癌细胞增殖的作用;利用4个数据库交叉比对和文献回顾,筛选得到5个感兴趣的潜在基因,包括细胞周期蛋白依赖性激酶抑制剂(ICK)、B细胞异位基因3(BTG3)、含CUB和LCCL结构域盘状蛋白2(DCBLD2)、真核起始因子4F(EIF4G2)和核不均一核糖核蛋白F(HNRNPF)。Western blot实验和双荧光素报告分析发现miR-139在肝细胞肝癌中具有直接抑制BTG3基因表达的作用。结论 miR-139可以直接调控BTG3基因的表达,影响肝癌细胞增殖。

Objective To determine the role of miR-139 in the development of hepatocellular carcinoma（ HCC） and to preliminarily clarify the underlying mechanism. Methods CCK-8 assay was performed to investigate the effect of miR-139 on HCC cell proliferation. Four databases,Target Scan,MiRanda,Clip-seq and miRDB,in combination with literature review,were used to predict the downstream pathways and specifically the potential targets of miR-139. The predicted targets were then verified by Western blot analysis and luciferase reporter assay. Results CCK-8 assay confirmed that miR-139 suppressed the proliferation of HCC cells. By the comparisons between the four databases and the review of literatures,a total of five genes（ ICK,BTG3,DCBLD2,EIF4 G2 and HNRNPF） were predicted as potential targets of miR-139. Western blot and luciferase reporter assay validated that miR-139 could directly inhibit the expression of BTG3 in HCC cells.Conclusion miR-139 can repress the proliferation of HCC cells via directly inhibiting the expression of BTG3.