miR-17-5p通过靶向作用AKT3抑制肝癌侵袭和迁移  

miR-17-5p inhibits invasion and metastasis of hepatocellular carcinoma through targeting effect on AKT3

作　　者：孔伟浩 李坤[1] 肖翠翠 胡竞雄 黄泽楠[3] 陈强星 张剑[1] Kong Weihao;Li Kun;Xiao Cuicui;Hu Jingxiong;Huang Zenan;Chen Qiangxing;Zhang Jian(Department of Liver Transplantation,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China;Department of Hepatobilliary Surgery,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China;Department of Thyroid and Breast Surgery,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院肝移植科,广州510630 [2]中山大学附属第三医院肝胆外科,广州510630 [3]中山大学附属第三医院甲乳外科,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2018年第2期152-155,共4页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：广东省科技计划项目(2014A020212159);广州市科技计划项目(201707010112)

摘　　要：目的探讨micro RNA(mi R)-17-5p对肝细胞癌(肝癌)细胞侵袭和迁移的影响及机制。方法采用RT-PCR检测L-02人正常肝细胞及QGY-7703肝癌细胞中mi R-17-5p的表达。mi R-17-5p模拟物和模拟物对照分别转染QGY-7703肝癌细胞。Transwell和划痕试验检测mi R-17-5p对肝癌细胞侵袭和迁移能力的影响。生物信息学分析确定mi R-17-5p的靶基因,Western blot检测其在肝癌细胞中表达。对靶基因行si RNA沉默后,观察肝癌细胞的侵袭和迁移能力。两种细胞中micro RNA比较采用t检验。结果 mi R-17-5p在肝癌细胞中表达量为0.16±0.04,较L-02正常肝细胞的1.01±0.19明显下调(t=-9.67,P<0.05)。转染mi R-17-5p模拟物的肝癌细胞穿膜细胞数、迁移速度分别为(36±4)个、(5.37±0.15)mm/d,明显低于对照组的(62±7)个、(7.50±0.01)mm/d(t=-15.40,-32.00;P<0.05)。生物信息学分析显示AKT3是mi R-17-5p的关键靶基因,AKT3在肝癌细胞中表达明显低于正常肝细胞。转染si RNA-AKT3的肝癌细胞穿膜细胞数、迁移速度分别为(13±3)个、(4.13±0.15)mm/d,明显低于对照组的(58±3)个、(7.23±0.25)mm/d(t=-17.88,-53.69;P<0.05)。结论 mi R-17-5p通过靶向作用于AKT3抑制肝癌细胞的侵袭和转移能力。Objective To explore the effect and mechanism of microRNA(miR)-17-5p on the invasion and metastasis of hepatocellular carcinoma(HCC)cells.Methods Expression of miR-17-5p in the normal human L-02 hepatocyte and QGY-7703 HCC cells was detected by RT-PCR.QGY-7703 HCC cells were transfected by miR-17-5p mimic and mimic control respectively.Influence of miR-17-5p on the invasion and metastasis ability of HCC cells was detected using Transwell assay and scratch test.Target gene of miR-17-5p was confirmed by bioinformatic analysis,and its expression in HCC cells was detected by Western blot.After siRNA silenced by target gene,the invasion and metastasis ability of HCC cells were observed.Comparison of microRNA in the two kinds of cells was conducted by t test.Results Expression level of miR-17-5p in HCC cells was 0.16±0.04,significantly lower than 1.01±0.19 in normal L-02 hepatocytes(t=-9.67,P<0.05).Number of trans-membrane cells and metastasis rate of HCC cells transfected by miR-17-5p mimic were respectively 36±4 and(5.37±0.15)mm/d,significantly lower than 62±7 and(7.50±0.01)mm/d of control group(t=-15.40,-32.00;P<0.05).Bioinformatic analysis showed that AKT3 was the key target gene of miR-17-5p,and the expression of AKT3 in HCC cells was obviously higher than that of normal hepatocyte.Number of trans-membrane cells and metastasis rate of HCC cells transfected by siRNA-AKT3 were respectively 13±3 and(4.13±0.15)mm/d,significantly lower than 58±3 and(7.23±0.25)mm/d of control group(t=-17.88,-53.69;P<0.05).Conclusion miR-17-5p inhibits the invasion and metastasis ability of HCC cells through targeting effect on AKT3.

关 键 词：癌 肝细胞 miR-17-5p AKT3 肿瘤侵润 细胞迁移 

分 类 号：R735.7[医药卫生—肿瘤]