摘要
目的研究外源性骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)过表达对骨肉瘤细胞的影响与Wnt/β-catenin信号通路的关系。方法用携带人BMP9基因的重组腺病毒Ad BMP9转染人骨肉瘤细胞MG63,上调MG63中BMP9的表达水平;MTT法检测细胞增殖能力的变化,Hochest染色法检测细胞凋亡的变化,Transwell实验检测细胞迁移能力的变化;实时定量PCR和Western blot分别检测Wnt/β-catenin信号通路中相关分子β-catenin、c-myc、cyclin D1和E-cadherin在mRNA和蛋白水平表达的变化。统计分析实验结果。结果获得了过表达BMP9的MG63细胞;BMP9能促进MG63细胞的增殖、迁移和凋亡;MG63细胞中BMP9的过表达增强了Wnt/β-catenin信号通路中相关分子β-catenin、c-myc、cyclin D1的表达,减弱了E-cadherin的表达。结论 BMP9对于骨肉瘤细胞MG63具有正性调节作用,该调控作用可能与Wnt/β-catenin信号通路相关。
Objective To investigate the effect of exogenous bone morphogenetic protein 9 （BMP9） on osteosarcoma cell line MG63 and its relationship with Wnt/β-catenin signaling pathway. Methods Recombinant adenovirus AdBMP9 carrying BMP9 gene was used to up-regulate the expression of BMP9 protein in MG63 cells. Then, MTr assay, Hoechst staining, and Transwell assay were used to detect the cell proliferation, apoptosis, and migration, respectively. The mRNA levels of genes involved in Wnt/β-catenin signaling pathway, including β-catenin, c-myc, cyclinD1 and E-cadherin, were examined by quantitative real-time PCR （qRT-PCR）, while the protein levels were determined by Western blotting. Results BMP9 protein was over-expressed in the MG63 cells after transfection of recombinant adenovirus. BMP9 could promote the proliferation, apoptosis and migration in the MG63 cells. The over-expression of BMP9 protein up-regulated the expressions of β-catenin, c-myc and cyclinD1, and down-regulated the expression of E-cadherin. Conclusion BMP9 can positively regulate the MG63 cells through Wnt/β-catenin signaling pathway.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2015年第7期655-659,共5页
Journal of Third Military Medical University
基金
甘肃省科技厅自然科学研究基金(1208RJZA272)~~
