Noggin基因过表达重组腺病毒载体的构建及其对骨肿瘤细胞MG63迁移侵袭能力的影响

Construction of Recombinant Adenovirus Vectors Overexpressing Noggin Gene and Its Effect on the Migration and Invasion Ability of Bone Tumor Cells MG63

目的:研究Noggin基因对骨肿瘤细胞MG63中Noggin,BMP-2,血管内皮生长因子(VEGF)及结缔组织生长因子(CTGF)的表达及细胞迁移侵袭的影响,探讨Noggin基因在骨肿瘤的发生和发展过程中的作用机制。方法:通过构建Noggin基因过表达重组腺病毒载体,并将其转染到骨肿瘤细胞MG63中,采用RT-PCR和Western blot分别检测空白对照组、过表达重组腺病毒组中Noggin,BMP-2,VEGF和CTGF的mRNA水平和蛋白水平,Transwell细胞迁移和侵袭能力实验考察两组细胞的迁移和侵袭能力。结果:成功构建Noggin基因过表达重组腺病毒载体,将其转染到骨肿瘤细胞MG63后,Noggin的mRNA及蛋白表达水平显著升高,BMP-2,VEGF及CTGF的mRNA及蛋白表达水平显著降低,转染Noggin基因过表达重组腺病毒的骨肿瘤细胞MG63迁移和侵袭的能力明显下降。结论:过表达Noggin基因能成功抑制骨肿瘤细胞MG63的侵袭和转移,有望成为骨肿瘤基因治疗的新方法。

Objective：To investigate the effect of Noggin gene on the expression of Noggin,BMP-2,vascular endothelial growth factor（VEGF）and connective tissue growth factor（CTGF）and the cell migration and invasion in bone tumor cells MG63,and to explore the mechanism of Noggin gene in the development and progression of bone tumors.Methods：The Noggin gene overexpressing recombinant adenovirus vectors were constructed and transfected into bone tumor cells MG63.The mRNA and protein levels of Noggin,BMP-2,VEGF,and CTGF in the blank control group and the overexpressing recombinant adenovirus group were detected by RT-PCR and Western blot respectively,and the migration and invasion ability of cells in the two groups were validated by Transwell cell migration and invasion ability experiment.Results：After the Noggin overexpressing recombinant adenovirus vectors were successfully constructed and transfected into bone tumor cells MG63,the mRNA and protein expression of Noggin were increased significantly,the mRNA and protein expression of BMP-2,VEGF and CTGF were inhibited significantly,and the migration and invasion ability of bone tumor cells MG63 infected with Noggin gene overexpressing recombinant adenovirus was decreased significantly.Conclusion：Overexpression of Noggin gene can inhibit the invasion and metastasis of bone tumor cells MG63 successfully,which is expected to be a new method for gene therapy of bone tumors.