葡萄糖转运蛋白5低表达对骨肉瘤MG-63细胞系克隆形成能力和迁移能力的影响及其可能机制

Effect of low expression of glucose transporter 5 on cloning and migration capacities of osteosarcoma MG-63 cell line and its possible mechanism

目的分析抑制葡萄糖转运蛋白5(Glut5)的表达对骨肉瘤MG-63细胞系克隆形成能力和迁移能力的影响,并基于Wnt/β-连环蛋白信号通路探讨其可能的作用机制。方法取对数生长期的MG-63细胞分为Con组、NC组和短发夹RNA(shRNA)-Glut5组。Con组未进行基因干扰,NC组和shRNA-Glut5组则采用RNA干扰技术将杂序的shRNA、shRNA-Glut5分别转染至MG-63细胞。转染48 h后,检测3组细胞的增殖能力、细胞克隆数和迁移能力,以及细胞中Glut5、β-连环蛋白、细胞周期蛋白D1(cyclin D1)和基质金属蛋白酶3(MMP3)的mRNA和蛋白的表达水平。结果与其他两组比较,shRNA-Glut5组MG-63细胞的增殖和迁移能力下降,细胞克隆数减少,Glut5、β-连环蛋白、cyclinD1和MMP3的mRNA和蛋白表达水平均降低(均P<0.05)。结论下调Glut5基因表达可以抑制骨肉瘤MG-63细胞系的增殖、克隆形成和迁移能力,Wnt/β-连环蛋白信号通路可能参与调控该过程。

Objective To analyze the effect of inhibiting expression of glucose transporter 5(Glut5)on cloning and migration capacities of osteosarcoma MG-63 cell line,and to explore its possible action mechanism based on Wnt/β-Catenin signaling pathway.Methods MG-63 cells at the logarithmic growth phase were divided into Con group,NC group and short hairpin RNA(shRNA)-Glut5 group.The Con group did not receive gene interference,in the NC and shRNA-Glut5 groups,RNA interference technique was used to perform transfection of scrambled shRNA and shRNA-Glut5 into MG-63 cells respectively.After 48 hours of transfection,cell proliferation capacity,number of cell cloning,and cell migration capacity,as well as the mRNA and protein expression levels of Glut5,β-Catenin,cyclin D1 and matrix metalloproteinase 3(MMP3)in MG-63 cells were determined in the three groups.Results Compared with the other two groups,the shRNA-Glut5 group yielded decreased proliferation capacity and migration capacity of MG-63 cells,exhibited decreased cell cloning number and lower mRNA and protein expression levels of Glut5,β-Catenin,cyclin D1 and MMP3(all P<0.05).Conclusion Down-regulation of Glut5 gene expression can inhibit the proliferation,cloning and migration capacities of osteosarcoma MG-63 cell line,and Wnt/β-Catenin signaling pathway may involve in regulating this process.