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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 (hBMP9) on human osteosarcoma cells MG63 and U2OS and its mechanism. Methods The recombinant adenovirus AdBMP9 was used to infect MG63 and U2OS cells. The blank control group (without any treatment factor) and the control group of AdGFP infection were also infected. ICC and Western blot were used to detect the expression of the two cell lines HBMP9 expression level; MTT and trypan blue exclusion of viable cell count method to detect cell proliferation activity; Hoechst / PI fluorescence double staining method to detect cell apoptosis; scratch healing test to detect cell migration ability; ICC test Expression of β-catenin in Wnt / β-catenin signaling pathway. Results The expression levels of hBMP9 in both AdBMP9-infected cells were significantly higher than those in blank control group and AdGFP-infected group (P <0.05). The upregulation of hBMP9 expression inhibited the proliferation of MG63 and U2OS cells in a time-dependent manner (P < 0.01). The apoptosis rate of both cell lines was significantly increased (P <0.01), the migration ability was significantly decreased (P <0.01), the expression of β-catenin was significantly decreased (P <0.01). Conclusion hBMP9 may inhibit the proliferation, migration and apoptosis of osteosarcoma cells by down-regulating Wnt / β-catenin signaling pathway.