To investigate the effect of Parkinson's disease related protein DJ-1 on the cell proliferation, apoptosis, invasion and migration in human osteosarcoma cells and the underlying molecular mechanisms. Methods: The protein expression levels of DJ-1 were detected in human osteosarcoma cell lines (MG-63, Saos-2, and U2OS) and human osteoblast cell line hFOB1.19 with or without deficiency in phosphatase and tensin homolog deleted from chromosome 10 (PTEN) were detected by Western blot. Osteosarcoma cells were treated with DJ-1 siRNA, and then the protein expression levels of DJ-1 were detected by Western blot. Cell survival rate of osteosarcoma cells was detected by cell counting kit-8 (CCK-8) assay. Cell apoptosis of osteosarcoma cells was measured by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining method. Cell invasive and migration ability of osteosarcoma cells were examined by transwell invasion and migration assay. Results: Compared with that of human osteoblast cell line (hFOB1.19), the protein expression level of DJ-1 was significantly upregulated in human osteosarcoma cell lines (MG-63, Saos-2, and U2OS) (all P<0.05), and U2OS had the highest level of DJ-1 when compared with the other three cell lines (P<0.01). DJ-1 siRNA could significantly down-regulate the DJ-1 protein expression in U2OS cells, and also diminish the cell survival rate. Moreover, DJ-1 down-regulation of DJ-1 could promote cell apoptosis, suppress the ability of cell invasion and migration, and increase the PTEN protein expression level (all P<0.05). In addition, the protein expression level of PTEN was markedly up-regulated in human osteosarcoma cell lines when compared with that in the hFOB1.19 cells (P<0.05). Conclusion: DJ-1 can promote the cell proliferation, inhibit cell apoptosis, and decrease the ability of cell invasion and migration, and the potential underlying mechanisms may be associated with the up-regulation of PTEN protein expression.
目的：研究帕金森病相关蛋白DJ-1对人骨肉瘤细胞增殖、凋亡、侵袭和迁移的影响及其作用机制。方法：Western 印迹检测骨肉瘤细胞株(MG-63，Saos-2和U2OS)及正常人成骨细胞株hFOB1.19中DJ-1和第10号染色体上缺失的磷酸酶与张力蛋白同源物(phosphatase and tensin homolog deleted from chromosome 10，PTEN)基因的表达。DJ-1 siRNA处理人骨肉瘤细胞后，采用Western印迹检测DJ-1的蛋白表达水平；细胞计数试剂盒(cell counting kit-8，CCK-8)法检测细胞存活率；膜联蛋白V(annexin V)-异硫氰酸荧光素(fluorescein isothiocyanate，FITC)/碘化吡啶(propidium iodide，PI)双染检测细胞凋亡；Transwell侵袭和迁移实验检测细胞侵袭和迁移水平。结果：与hFOB1.19细胞比较，骨肉瘤细胞DJ-1蛋白表达水平显著升高(均P<0.05)，其中U2OS细胞升高最为显著(P<0.01)。DJ-1 siRNA可显著降低U2OS细胞中DJ-1蛋白表达水平、降低细胞存活率、促进细胞凋亡、抑制细胞侵袭和迁移水平，以及增加PTEN蛋白表达水平(均P<0.05)。另外，与hFOB1.19细胞比较，PTEN在骨肉瘤细胞中的蛋白表达水平显著降低(P<0.05)。结论：DJ-1可抑制人骨肉瘤细胞增殖，促进细胞凋亡，抑制细胞侵袭和迁移水平，其机制可能与增加PTEN蛋白的表达水平有关。.