miR-370及其靶基因对肝癌细胞转移的影响及作用机制  

Effect of miR-370 and Its Target Genes on Metastasis of Hepatocellular Carcinoma Cells and Its Mechanism

作　　者：黄晓峰[1] 张翔[1] 唐波[1] 王宇笛 Huang Xiaofeng;Zhang Xiang;Tang Bo;Wang Yudi(PLA 44 Hospital in Guiyang City,Guiyang 550009,China)

机构地区：[1]中国人民解放军第四十四医院,贵州贵阳550009

出　　处：《北华大学学报（自然科学版）》2018年第6期750-753,共4页Journal of Beihua University（Natural Science）

基　　金：贵州省科技厅重大专项课题(20142012-3)

摘　　要：目的探讨miR-370及其靶基因对肝癌细胞转移的影响及作用机制.方法通过脂质体2000分别将pc DNA-miR-370,pc DNA3.0转入HepG2细胞,建立miR-370过表达组、阴性对照组,进行Transwell迁移和侵袭实验、CCK8细胞增殖活性检测实验;应用流式细胞术分析细胞周期;应用miRanda、Targetscan数据库预测miR-370的可能靶基因Aeg-1,并通过双荧光素酶检测验证;应用Western blot检测Aeg-1蛋白表达情况.结果 miR-370过表达组HepG2细胞迁移能力和侵袭能力均明显低于阴性对照组,差异具有统计学意义(P<0.01); CCK8法检测miR-370过表达组细胞吸光度值明显低于阴性对照组,差异具有统计学意义(P<0.05); miR-370过表达组G0/G1期细胞比例明显高于阴性对照组,G2/M期细胞比例明显低于阴性对照组,差异具有统计学意义(P<0.05);miR-370与Aeg-1基因3'UTR载体共同转染Hep G2细胞后荧光素蛋白表达量明显降低,Aeg-1 3'UTR突变后荧光素蛋白表达量恢复; Western blot检测显示:miR-370过表达组Aeg-1蛋白的相对表达量明显低于阴性对照组,差异具有统计学意义(P<0.05).结论 miR-370可通过负调控其靶基因Aeg-1来抑制肝癌细胞转移的多个步骤,包括增殖、迁移、侵袭.Objective To investigate the effect of miR-370 and its target genes on metastasis of hepatocellularcarcinoma cells and its mechanism. Method pcDNA-miR-370 and pcDNA3. 0 were transferred into HepG2 cellsby liposome 2000 to establish miR-370 over-expression group and negative control group. The Transwell migrationand invasion experiment,the proliferation activity of CCK8 cells were detected,and the cell cycle was analyzed byflow cytometry. miRanda and Targetscan database were used to predict the possible target gene Aeg-1 of miR-370,double luciferase test and Western blot were used to detect the expression of Aeg-1 protein. Results Themigration and invasion ability of HepG2 cells in miR-370 over-expression group were significantly lower thanthose in negative control group （ P 〈 0. 01 ）. The cell absorbency of miR-370 over-expression group wassignificantly lower than that of negative control group （P〈0. 05）,and the proportion of G0 / G1 stage cells inmiR-370 over-expression group was significantly higher than that of negative control group （P〈0. 05）,and theproportion of G2 / M phase cells was significantly lower than that of negative control group,and the difference hasthe significance of overall planning （P〈0. 05）. The expression of fluorescein protein was significantly decreasedafter the transfection of miR-370 and Aeg-1 gene 3 ’UTR vector to HepG2 cells,and the expression of fluoresceinprotein was recovered after the mutation of Aeg-1 3’ UTR. Western blot test showed that the relative expression ofAeg-1 protein in the miR-370 over-expression group was significantly lower than that in the negative control group（P〈0. 05）. Conclusion miR-370 can inhibit the metastasis of hepatocellular carcinoma by regulating its targetgene Aeg-1,including proliferation,migration and invasion.

关 键 词：肝癌 转移 miR-370 AEG-1 

分 类 号：R735.7[医药卫生—肿瘤]