目的：探讨Grb2协同结合蛋白2（ Gab2）在人骨肉瘤细胞系中的表达及其与人骨肉瘤细胞侵袭转移的关系。方法：应用小RNA干扰技术，将合成的小RNA干扰质粒转染给人骨肉瘤U2-OS细胞，采用Western blotting和RT-PCR法检测转染后Gab2的蛋白和mRNA表达水平；体外细胞趋化运动实验和侵袭实验检测细胞的迁移和侵袭能力。结果：Gab2在人骨肉瘤U2-OS细胞系中高表达，且转染Gab2 siRNA后的细胞（ SiGab2／U2-OS）中Gab2蛋白及mRNA的表达水平低于转染scramble siRNA细胞（ Scr／U2-OS）和U2-OS；趋化运动实验发现在10μg／L表皮生长因子（ EGF）的诱导下SiGab2／U2-OS细胞的迁移能力较U2-OS和Scr／U2-OS细胞明显下降，差异有统计学意义（P＜0.01）；SiGab2／U2-OS细胞侵袭并穿透Matrigel 膜基质的细胞数量均比Scr／U2-OS细胞和U2-OS细胞少，SiGab2／U2-OS细胞的体外侵袭能力显著降低（P＜0.01）。结论：利用siRNA干扰技术降低Gab2的表达对人骨肉瘤U2-OS细胞的迁移和侵袭能力有明显的抑制作用。%AIM:To investigate the expression of Grb2-associated binding protein 2 (Gab2) in human osteo-sarcoma cells and its relationship with the invasion and metastases of human osteosarcoma cells.METHODS: The tech-nique of small RNA interference was used to transfect human osteosarcoma U2-OS cell lines.Western blotting and RT-PCR were used to detect the protein and mRNA expression of Gab2 in transfected U2-OS cells.After transfection, through chem-otaxis and invasion assays in vitro, the cell migration and invasion abilities were detected.RESULTS:After transfection, the expression of Gab2 at mRNA and protein levels in Gab2 siRNA transfected cells ( SiGab2/U2-OS) was lower than that in scrambled siRNA transfected cells ( Scr/U2-OS ) and U2-OS cells.After stimulation with epidermal growth factor ( EGF) at concentration of 10 μg/L, the migration SiGab2/U2-OS cells was significantly less than Scr/U2-OS cells and U2-OS cells ( P<0.01 ) .The number of invasion cells of SiGab2/U2-OS group was significantly lower than the other 2 control groups ( P<0.01) .CONCLUSION:Inhibition of Gab2 expression obviously attenuates the migration and invasion abilities of human osteosarcoma U2-OS cell line.
展开▼
