miR-129-5p靶向调控VCP抑制骨肉瘤细胞体外迁徙侵袭  

miR-129-5p suppresses osteosarcoma cells migration and invasion via targeting VCP in vitro

作　　者：周云飞[1] 龙新华[1] 张志宏[1] 陈宣银[1] 刘家明[1] 黄山虎[1] 刘志礼[1] 

机构地区：[1]南昌大学第一附属医院骨科,南昌330006

出　　处：《临床与实验病理学杂志》2014年第10期1139-1143,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：国家自然科学基金(81360399);江西省自然基金(2012ZBAB205016)

摘　　要：目的探讨miR-129-5p是否靶向调控VCP基因抑制骨肉瘤细胞迁徙侵袭。方法构建miR-129-5p过表达及低表达的慢病毒载体,转染骨肉瘤细胞U2-OS;采用实时荧光定量PCR检测上调及下调miR-129-5p的U2-OS细胞中miR-129-5p的表达量;采用RT-PCR和Western blot技术检测VCP mRNA和蛋白表达;采用划痕实验和Transwell侵袭实验检测细胞迁徙、侵袭情况。结果实时荧光定量PCR结果显示U2-OS细胞中miR-129-5p表达被明显上调或下调;RT-PCR和Western blot检测结果显示:miR-129-5p上调U2-OS细胞组中VCP mRNA和蛋白表达水平显著低于阴性对照细胞组(阴性慢病毒转染);miR-129-5p下调U2-OS细胞组中VCP mRNA和蛋白表达水平显著高于阴性对照细胞组;miR-129-5p上调U2-OS细胞迁徙和侵袭力显著低于阴性对照细胞,miR-129-5p下调的U2-OS细胞迁徙和侵袭力显著高于阴性对照细胞。结论 miR-129-5p靶向调控VCP的表达而抑制骨肉瘤细胞迁徙和侵袭能力。Purpose miR-129-5p was demonstrated to be one of genes targeting VCP by luciferase reporter assay previously. This study was to investigate the effect of miR-129-5p on cell invasion and migration of U2-OS cell by targeting to VCP in vitro. Methods Lentivirus vectors were constructed to up-regulate or down-regulate the expression of miR-129-Sp in human osteosareoma cell U2-OS. The expression of miR-129-5p in up-regulated and down-regulated U2-OS cells was assessed by quantitative real time PCR. The VCP mRNA and protein expression was assessed by RT-PCR and Western blot, respectively. Cell invasion and migration ability were detec- ted by transwell invasion assay and wound-healing assay. Results RT-PCR and Western blot analysis testified that both the expression of VCP mRNA and protein were significantly decreased in miR-129-5p up-regulated U2-OS cells, compared to cells in miR-129-5p down-regulated group and negative control group. Remarkable reduction of cell migration and invasion ability was observed in miR-129- 5p up-regulated ceils, but it was on the contrary in miR-129-5p down-regulated cells. Conclusions miR-129-5p can inhibit the mi- gration and invasion of human osteosarcoma cells via targeting VCP.

关 键 词：骨肉瘤 迁移 侵袭 miR-129-5p VCP 

分 类 号：R738.1[医药卫生—肿瘤]