[摘要] 目的:探讨葡萄糖转运蛋白-1（glucose transporter-1，Glut-1）通过Wnt/β-catenin 通路促进骨肉瘤细胞MG63 的迁移及其机制。方法: 构建针对Glut-1 基因的RNA 干扰重组腺病毒（Ad-Glut-siRNA）及含对照序列的重组腺病毒（Ad-GFP），感染MG63 细胞沉默Glut-1 基因表达；通过Transwell小室法检测Blank组、Ad-AFP组、Ad-Glut-siRNA 组、GSK-3 抑制剂AZD2858 组细胞的迁移能力；应用免疫荧光实验检测各组细胞E-钙黏蛋白（E-cadherin）、波形蛋白（vimentin）的表达变化以及β-连环蛋白（β-catenin）的进核情况，通过Western blotting 实验检测各组细胞的MMP-2、MMP-9 表达变化以及Blank 组、Ad-AFP组、Ad-Glut-siRNA组细胞FZD7、β-catenin、Dsh 蛋白表达变化。结果: 沉默Glut-1 基因后，MG63 细胞迁移能力明显下降（P<0.05）；再给于AZD2858 处理后，细胞的迁移能力恢复（P<0.05）。与Blank 组和Ad-GFP组细胞相比，Ad-Glut-siRNA 组MG63 细胞E-cadherin 表达显著升高（P<0.05），vimentin、MMP-2、MMP-9 以及FZD7、β-catenin、Dsh蛋白表达显著降低（均P<0.05）；与Ad-Glut-siRNA组相比，AZD2858 组细胞E-cadherin 表达显著降低（P<0.05），vimentin、MMP-2 和MMP-9 表达显著升高（P<0.05）。结论: Glut-1 基因高表达与骨肉瘤MG63 细胞侵袭转移具有密切联系，其可能机制在于Glut-1 高表达后激活Wnt/β-catenin 通路使EMT相关蛋白Ecadherin表达降低、vimentin以及MMP-2、MMP-9含量上升，从而促进MG63 细胞的转移。

[Abstract] Objective: To explore the mechanism of glucose transport protein-1(Glut-1) promoting the migration of osteosarcoma MG63 cells through Wnt/β-catenin pathway. Methods: RNA interference recombinant adenovirus targeting Glut-1 gene (Ad-Glut-siRNA)and control recombinant adenovirus (Ad-GFP) were constructed and transfected into MG63 cells to silence Glut-1 gene expression.The cell migration ability of Blank group, Ad-AFP group, Ad-Glut-siRNA group and AZD2858 (inhibitor of GSK-3) group were detected by Transwell chamber migration assay. Immunofluorescence assay was used to detect the expression of E-cadherin and vimentin in each group and the nuclear translocation of β-catenin. The expression of MMP-2 and MMP-9 in each group and FZD7, β-catenin,Dsh protein in Blank group, Ad-AFP group, Ad-Glut-siRNA group were detected by Western blotting assay. Results: The migration ability of MG63 cells was significantly decreased (P<0.05) after Glut-1 gene silencing, which was restored after AZD2858 treatment (P<0.05). Compared with Blank group and Ad-GFP group, the E-cadherin level in MG63 cells in Ad-Glut-siRNA group was significantly increased (P<0.05), while the expressions of vimentin, MMP-2, MMP-9, FZD7, β-catenin and Dsh protein were significantly reduced (all P<0.05). Compared with Ad-Glut-siRNA group, E-cadherin expression of AZD2858 group was significantly reduced, while the expressions of vimentin, MMP-2, MMP-9 were significantly up-regulated (P<0.05). Conclusion: The high expression of Glut-1 gene is closely related to the invasion and metastasis of MG63 cells. The possible mechanism is that the high expression of Glut-1 leads to the activation of Wnt/β-catenin pathway, which leads to the decrease of EMT-related protein E-cadherin, and the increase of vimentin and MMP-2, MMP-9, and further promotes the migration of MG63 cells.

国家自然科学基金资助项目（No. 81371230）