上调LRIG1对胶质瘤U251细胞侵袭、迁移的影响

目的 探讨上调LRIG1表达对胶质瘤U251细胞侵袭及迁移的影响。方法 体外培养U251细胞，应用LipofectamineTM 2000试剂盒将质粒pLRIG1-GFP（pLRIG1-GFP组）及其空载pEGFP-N1质粒（pEGFP-N1组）转染U251细胞，G418挑选具有抗性的细胞并扩增，以供进一步实验。另选择不转染任何质粒U251细胞作为空白对照组。qRT-PCR、免疫印迹法检测LRIG1及细胞上皮-间充质转化（EMT）标志蛋白SNAI2、E-cadherin、N-cadherin和vimentin的mRNA和蛋白表达；应用Transwell小室实验检测U251细胞侵袭和迁移能力。结果 成功构建高表达LRIG1的胶质瘤U251细胞，荧光显微镜下可见细胞散发绿色荧光。pLRIG1-GFP组LRIG1 mRNA及蛋白表达水平明显高于pEGFP-N1组和空白对照组（P<0.05），而后两组之间均无统计学差异（P>0.05）。pLRIG1-GFP组侵袭和迁移细胞数明显低于pEGFP-N1组和空白对照组（P<0.05），而后两组之间均无统计学差异（P>0.05）。pLRIG1-GFP组SNAI2、N-cadherin、vimentin mRNA及蛋白表达水平明显低于pEGFP-N1组和空白对照组（P<0.05），而E-cadherin mRNA和蛋白表达水平明显高于pEGFP-N1组和空白对照组（P<0.05）；后两组之间均无统计学差异（P>0.05）。结论 上调LRIG1，抑制U251细胞侵袭和迁移，其机制可能与调控细胞EMT过程有关。

Effect of up-regulation of LRIG1 on invasion and migration of glioma U251 cells

Objective To explore the effect of up-regulation of leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) on the invasion and migration of gliona U251 cells. Methods The U251 cells were cultured in vitro and than transfected with plasmids pLRIG1-GFP (pLRIG1-GFP group) and pEGFP-N1 (pEGFP-N1 group). The U251 cells which were risitant to G418 were used for further experiments. Those cells without transfection of pLRIG1-GFP or pEGFP-N1 were used as blank control group. The mRNA and protein expressions of LRIG1, and biomarkers of epithelial-to-mesenchymal transition (EMT) including SNAI2, E-cadherin, N-cadherin and vimentin were detected using qRT-PCR and western blot, respectively. The invasion and migration abilities of U251 cells were detected by Transwell experiments. Results The glioma U251 cells with high expression of LRIG1 were successfully constructed. The expression levels of LRIG1 mRNA and protein in the pLRIG1-GFP group were significantly higher than those in the pEGFP-N1 group and the blank control group (P<0.05), but there was no statistical difference between groups pEGFP-N1 and blank control (P>0.05). The number of invasion and migration cells in the pLRIG1-GFP group was significantly lower than that in the pEGFP-N1 group and the blank control group (P<0.05), but there was no statistical difference between groups pEGFP-N1 and blank control (P>0.05). The mRNA and protein expression levels of SNAI2, N-cadherin and vimentin in the pLRIG1-GFP group were significantly lower than those in the pEGFP-N1 group and the blank control group (P<0.05), the mRNA and protein expression levels of E-cadherin were significantly higher than those in the pEGFP-N1 group and blank control group (P<0.05); there was no statistical difference between groups pEGFP-N1 and blank control (P>0.05). Conclusion Up-regulation of LRIG1 can inhibit the invasion and migration of U251 cells, which may be related to the regulation of cell EMT process.