miRNA沉默VCP对骨肉瘤细胞U2-OS增殖和迁移的影响  

Effects of miRNA-mediated VCP gene silencing on the proliferation and migration of human osteosarcoma cell line U2-OS in vitro

作　　者：龙新华[1] 刘志礼[1] 黄山虎[1] 毛建华[1] 王高[2] 周扬[1] 

机构地区：[1]南昌大学第一附属医院骨科,南昌330006 [2]南昌大学第三附属医院骨科,南昌330006

出　　处：《广东医学》2013年第10期1481-1485,共5页Guangdong Medical Journal

基　　金：江西省教育厅科学技术研究项目(编号:GJJ11316)

摘　　要：目的探讨沉默VCP基因对人骨肉瘤细胞系增殖、迁移的影响。方法构建靶向VCP基因的重组质粒载体(X135-1-2、X135-2-1、X135-3-1)和阴性对照载体(Negative),测序鉴定;X135-1-2、X135-2-1、X135-3-1分别转染人骨肉瘤细胞系U2-OS(阳性干扰组:X135-1-2组、X135-2-1组、X135-3-1组),Negative质粒转染为阴性对照组,未转染细胞为空白对照组。RT-PCR检测VCP mRNA的表达水平,Western blot测定VCP的蛋白表达水平,MTT检测细胞的增殖情况,Transwell migration实验检测细胞迁移力。结果转染后48h,与空白对照组和阴性对照组比较,X135-1-2、X135-2-1和X135-3-1组VCP mRNA和蛋白的表达均明显降低,尤以X135-3-1组更为显著[VCP mRNA和蛋白表达抑制效率分别为(81.4±3.0)%和(83.3±6.0)%];各阳性干扰组细胞的增殖能力均被明显抑制,以X135-3-1组更为显著(P<0.05);各阳性干扰组细胞的迁移能力明显下降,以X135-3-1组更为显著(P<0.05)。结论沉默VCP能有效抑制人骨肉瘤细胞系U2-OS细胞的增殖和迁移,VCP可能是骨肉瘤治疗的分子靶点。Objective To investigate the effects of VCP knockdown on the proliferation and migration of human osteosarcoma cell line in vitro. Methods Three recombinant plasmids （X135 - 1 - 2, X135 - 2 - 1 and X135 - 3 - 1, respectively） targeting VCP gene were constructed, so was a negative plasmid as negative control; which were identified and transfected to human osteosarcoma cell line. The normal cell line was selected as positive control. The VCP mRNA and protein expression were assessed by PCR and western blot, respectively; whereas MTT assay and transwell migration assay were carried out for was performed for assessment of cell proliferation and migration ability. Results There were significant down -regulation in both mRNA and protein of VCP 48 hours after transfection, which were by 81.4 ± 3.0% and 83.3 ±6. 0%, respectively, in X135 -3 -1 group. The cell proliferation was significantly inhibited in each VCP - knockdown group （P 〈 0. 05 ）, so was the migration ability （P 〈 0. 05） ; which were significantly most prominent in X135 - 3 - 1 group （ P 〈 0. 05 ）. Conclusions VCP gene silencing efficiently inhibits the proliferation and migration of U2 - OS cells in vitro, suggesting a potential molecular target for osteosarcoma therapy.

关 键 词：基因沉默 VCP 骨肉瘤 增殖 迁移 

分 类 号：R730.3[医药卫生—肿瘤]