mir-520c-3p靶向GPC3对肝癌Huh-7细胞增生、迁移和侵袭能力的影响的研究  

Mir-520c-3p targeted GPC3 for hepatocellular carcinoma Huh-7cell proliferation, migration and invasion ability

作　　者：刘忠考[1] 雷长江[2] 邱志东[2] 李明意[2] 黎然[1] 包仕廷[1] 缪辉来[2] 

机构地区：[1]广东医学院附属医院肝胆外科,湛江524001 [2]广东医学院附属医院肝胆外科研究室,湛江524001

出　　处：《国际外科学杂志》2013年第4期238-242,F0003,共6页International Journal of Surgery

基　　金：基金项目：广东省科技计划项目（No.20101700501）

摘　　要：目的研究mir-520c-3p靶向GPC3后对肝癌Huh-7细胞增生、迁移、侵袭能力的影响，为临床寻找治疗肝癌的方法提供新的理论依据。方法将实验分为三组：未转染mir-520c-3p的Huh-7细胞组（cell组），阴性对照组（NC组），以及转染mir-520c-30p的Huh-7细胞组（干扰组）。然后利用荧光定量PCR和WesternBlotting分别检测GPC3基因的mRNA和蛋白的表达量；通过EDU检测细胞增生的变化；通过Transwell检测细胞侵袭和迁移能力的改变。结果荧光定量PCR结果表明cell组、NC组和干扰组分别为1．13±0．23、1．28±0．15和1．05±0．19（P〉0．05），mir-520e-3p不影响GPC3的mRNA转录水平；而West—ernBlotting实验结果表明转染mir-520c-3p后GPC3蛋白表达水平明显降低，cell组、NC组和干扰组分别为2．16±0．08、1．99±0．04和0．499±0．05（P〈0．01）；EDU检测发现转染mir-520c-3p后肝癌Huh-7细胞增生能力明显受抑制，cell组、NC组和干扰组分别为（90．12±1．93）％、（91．02±0．35）％和（77．73±5．88）％（P〈0．05）；Transwell检测结果发现转mir-520c-3p后肝癌Huh-7细胞的侵袭能力受抑制，cell组、NC组和干扰组分别为0．071±0．008、0．105±0．001和0．048±0．002（P〈0．05）；迁移能力明显降低，其中cell组、NC组和干扰组分别为0．5464-0．010、0．328±0．002和0．151±0．002（P〈0．01）。结论mir-520c-3p能够靶向GPC3从而抑制肝癌Huh-7细胞的增生、侵袭、迁移能力。Objective To research the effect of mir-520c-3p targeted GPC3 to the hepatocellular carcinoma Huh-7 cell proliferation, migration, and the influence of the attack ability and find new theoretical basis for liver hepatocellular carcinoma clinical treatment. Methods The cells were divided into three groups： not transfection of mir-520c-3p group （ cell group） , negative control group （ Nc group） , and transfection of mir-520c-3p group （ treat ment group）. Then used fluorescence quantitative PCR and Western Blot to detect GPC3mRNA gene and protein expression quantity. Cell proliferation of change was detected by the EDU. Made use of Transwell to detect cell inva- sion and migration ability of the change. Results Fluorescence quantitative PCR results showed that Cell group, NC group and treatment group were 1.13 ± 0.23, 1.28 ±0.15 and 1.05 ± 0.19 （ P 〉 0.05 ）, mir- 520- 3p could not reduce the GPC3mRNA; but Western Blot detection results showed that GPC3 protein expression level reduce sig- nificantly after transfection mir-520c-3p, Cell, NC and treatment group were 2. 16 ± 0.08, 1.99±0.04 and 0.499 ±0.05 （P 〈 0. 01 ）. The EDU detection results showed that hepatoeellular carcinoma Huh-7 cell proliferation ability obviously inhibited after transfection mir-520c-3p, Cell group, NC group and treatment group were （90.12 ± 1.93）% , （91.02 ±0.35）% and （77.73 ±5.88）% （P 〈0.05）, and Transwell test found that hepato- cellular carcinoma Huh-7cell invasion abilities were restrained, Cell group, NC group and treatment group were 0. 071 ± 0. 008, 0. 105 ± 0. 001 and 0. 048 ±0. 002 （P 〈 0.05 ）, in the same the cells＇ migration abilities werereduced, Cell group, NC group and treatment group were 0.546 ±0. 010, 0.328 ± 0. 002 and 0.151 ±0. 002 （ P 〈 0.01 ）. Conclusions Mir-520c-3p can target GPC3 so that affect hepatocellular carcinoma Huh-7 cell prolifera- tion, invasion and migration abilities.

关 键 词：mir-520c-3p 肝细胞癌 GPC3基因 细胞增生 

分 类 号：R735[医药卫生—肿瘤]