人骨形态发生蛋白9对人骨肉瘤细胞MG63和U2OS的抑制作用及其机制  

Inhibitory effect of human bone morphogenetic protein 9 on human osteosarcoma MG63 and U2OS cells and relevant mechanism

作　　者：吴丽美[1] 叶立伟[1] 武睿[1] 段亮[1] 张昀源[1] 陈娴[1] 王海燕[1] 周兰[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国生物制品学杂志》2013年第5期664-669,共6页Chinese Journal of Biologicals

摘　　要：目的探讨人骨形态发生蛋白9(Human bone morphogenetic protein 9,hBMP9)对人骨肉瘤细胞MG63和U2OS的抑制作用及其机制。方法用重组腺病毒AdBMP9分别感染MG63和U2OS细胞,并设空白对照组(不加任何处理因素)和AdGFP感染对照组,免疫细胞化学法(Immunocytochemistry,ICC)和Western blot法检测感染后两种细胞中hBMP9的表达水平;MTT和台盼蓝拒染活细胞计数法检测细胞的增殖活力;Hoechst/PI荧光双染法检测细胞的凋亡情况;划痕愈合试验检测细胞的迁移能力;ICC法检测Wnt/β-catenin信号途径中β-catenin的表达。结果AdBMP9感染的两种细胞中hBMP9的表达水平均明显高于空白对照组和AdGFP感染组(P<0.05);hBMP9表达的上调可抑制MG63和U2OS细胞的增殖,且呈时间依赖性(P<0.01),并使两种细胞的凋亡率明显增加(P<0.01),迁移能力明显下降(P<0.01),β-catenin的表达量明显减少(P<0.01)。结论 hBMP9可能通过下调Wnt/β-catenin信号途径活性,抑制骨肉瘤细胞的增殖、迁移,并促进其凋亡。Objective To investigate the inhibitory effect of human bone morphogenetic protein 9（BMP9）on human osteosarcoma MG63 and U2OS cells as well as the relevant mechanism.Methods MG63 and U2OS cells were infected with recombinant adenovirus AdBMP-9,using those uninfected as blank control and those infected with AdGFP as parallel control,then determined for expression of hBMP9 by immunocytochemistry（ICC）and Western blot,for proliferative activity by MTT method and viable counting with trypan blue exlusion method,for apoptosis by Hoechst/PI fluorescent staining,for migration ability by wound healing test,and for expression of β-catenin in Wnt/β-catenin signaling pathway by ICC assay.Results The expression levels of hBMP9 in MG63 and U2OS cells infected with AdBMP9 were significantly higher than those in blank and parallel control groups（P〈0.05）.The up-regulation of hBMP9 expression showed time-dependent inhibitory effect on the proliferations of MG63 and U2OS cells（P〈0.01）,while increased the apoptosis rate（P〈0.01）and decreased the migration ability（P〈0.01）and β-catenin expression（P〈0.01）of the cells.Conclusion The hBMP9 might inhibit the proliferation and migration while promote the apoptosis of osteosarcoma cells through Wnt/β-catenin signaling pathway.

关 键 词：骨形态发生蛋白质 骨肉瘤 细胞增殖 细胞迁移 细胞凋亡 Β-CATENIN 

分 类 号：R738.1[医药卫生—肿瘤]