葡萄糖转运蛋白-1通过Wnt/β-catenin通路促进骨肉瘤MG63细胞迁移  

Glucose transporter-1 promotes migration of osteosarcoma MG63 cells via the Wnt/β-catenin pathway

作　　者：郝俊龙[1] 王亚鹏 杨凯[1] 齐进[1] 王旭[1] 汪静[1] HAO Junlong, WANG Yapeng, YANG Kai, QI Jin, WANG Xun, WANG Jing(Department of Bone Oncology, The Second Affiliated Hospital of Lanzhou University, Lanzhou 730000, Gansu, Chin)

机构地区：[1]兰州大学第二医院骨肿瘤科,甘肃兰州730000

出　　处：《中国肿瘤生物治疗杂志》2018年第5期469-474,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81371230)~~

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-si RNA)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-si RNA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-si RNA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-si RNA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-si RNA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。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-si RNA） 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-si RNA 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-si RNA 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-si RNA 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-si RNA 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.

关 键 词：葡萄糖转运蛋白-1 骨肉瘤 MG63细胞 WNT/Β-CATENIN通路 迁移 

分 类 号：R738.1[医药卫生—肿瘤] R730.23[医药卫生—临床医学]