miRNA-224通过HOXD10影响Hep3B细胞迁移  

MiR-224 promotes cell migration by targeting HOXD10 in Hep3B cells

作　　者：丁琛琛 李琼[2] 王阁[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所肿瘤中心,重庆400042 [2]第三军医大学基础医学部教学实验中心,重庆400038

出　　处：《第三军医大学学报》2014年第13期1365-1369,共5页Journal of Third Military Medical University

基　　金：重庆市自然科学基金支持(CSTC2011jja0634)~~

摘　　要：目的探讨miR-224影响肝癌细胞迁移的可能机制。方法采用双荧光素酶实验、载体构建和Western blot实验等对预测的miR-224靶基因进行验证。qRT-PCR检测正常肝细胞株L02和肝癌细胞株Hep3B中miR-224的表达；划痕实验观察miR-224对肝癌细胞迁移的影响。结果qRT-PCR结果显示，与正常肝细胞株L02比较，miR-224在肝癌细胞株Hep3B中明显高表达（相对表达倍数1.679 2，P〈0.05）；划痕实验显示，与miR-224 mimics阴性对照组（迁移率：56.43%~58.33%）以及空白组（迁移率：60.48%~66.94%）比较， miR-224 mimics组（迁移率：80.12%~82.02%）的Hep3B细胞迁移能力明显增强（P〈0.05），反之亦然，提示miR-224表达水平和肝癌细胞的迁移能力成正相关；双荧光素酶实验结果显示：与对照组比较， miR-224 mimics组细胞的荧光信号明显下降（P〈0.05）；Western blot实验结果显示：与对照组比较，上调miR-224的表达水平后HOXD10表达下降，下调其表达水平HOXD10表达上升（P〈0.05）。结论 miR-224通过调控靶基因HOXD10的表达，参与调节肝癌细胞的迁移。Objective To investigate the possible underlying mechanism that miR-224 affects the migration of liver cancer cells. Methods A luciferase reporter assay was used to confirm that the gene HOXD10 was one direct target of miR-224. qRT-PCR, Western blotting, and scratch wound migration assay were used to clarify the molecular mechanism of miR-224 in the regulation of cell migration in human hepatocellular carcinoma line Hep3B. Results qRT-PCR indicated the expression of miR-224 was significantly stronger in Hep3B cells than in human hepatic L02 cells （relative expression ratio： 1.679 2, P〈0.05）. Scratch wound migration assay showed that transfection of miR-224 resulted in significantly increased ratio of migration （80.12% to 82.02%, P〈0.05） in Hep3B cells than the cells transfected by miR-224 mimic negative control （56.43% to 58.33%） and by blank vector （60.48% to 66.94%）. Compared with the miR-224 negative control, the luciferase activity was significantly decreased in 293 cells with miR-224 mimic （0.633 2）. miR-224 affected the migration ability of HCC via directly targeting HOXD10. Conclusion miR-224 is involved in the regulation of hepatoma cell migration via directly targeting HOXD10.

关 键 词：肝细胞肝癌 miRNA-224 HOXD10 迁移 

分 类 号：R394.3[医药卫生—医学遗传学] R73-37[医药卫生—基础医学]