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目的观察microRNA-1(miR-1)对肝癌细胞系Hep3B死亡结构域沉默子(BAG4)的表达及细胞凋亡的影响。方法 Western blot检测肝癌组织中BAG4蛋白表达情况;将过表达miR-1的质粒转染肝癌细胞Hep3B,分别用RT-qPCR及Western blot检测BAG4 mRNA和蛋白的表达,流式细胞仪检测细胞凋亡;生物信息学软件分析miR-1和BAG4 3’UTR作用关系。结果 BAG4蛋白在肝癌组织中的表达明显高于癌旁组织;Hep3B转染miR-1过表达质粒后,相较于阴性对照(NC)组,BAG4mRNA和蛋白水平的表达均有所降低;早期凋亡率明显增高。生物信息学分析提示BAG4是miR-1潜在靶基因。结论 miR-1可以诱导肝癌细胞Hep3B凋亡,其作用可能和抑制BAG4表达有关。
Objective To observe the effect of microRNA-1 (miR-1) on the expression of apoptosis-related gene Bcl-4 and its apoptosis in Hep3B hepatocellular carcinoma cell line Hep3B. Methods Western blot was used to detect the expression of BAG4 protein in hepatocellular carcinoma. The plasmid overexpression of miR-1 was transfected into Hep3B hepatocellular carcinoma cells. The expression of BAG4 mRNA and protein was detected by RT-qPCR and Western blot respectively. The apoptosis of hepatocellular carcinoma cell was detected by flow cytometry Bioinformatics software analyzed the interaction between miR-1 and BAG4 3'UTR. Results The expression of BAG4 protein in HCC tissues was significantly higher than that in adjacent non-cancerous tissues. After Hep3B transfection with miR-1 overexpression plasmid, the expression of BAG4 mRNA and protein decreased compared with the negative control group (NC) Mortality increased significantly. Bioinformatics analysis suggests that BAG4 is a potential target gene of miR-1. Conclusion miR-1 can induce apoptosis of Hep3B hepatocarcinoma cells, which may be related to the inhibition of BAG4 expression.