Mig-7通过MEK/ERK信号通路抑制胶质瘤U251侵袭及血管生成拟态

目的研究迁移诱导基因7（Mig-7）通过MEK/ERK信号通路抑制人脑胶质瘤细胞株U251体外血管生成拟态（VM）形成能 力和迁移、侵袭能力的影响。方法采用RNA干扰技术将特异性针对Mig-7基因的sh-Mig-7 转入人脑胶质瘤U251细胞中，并 观察感染效率；用携带有sh-Mig-7和阴性对照（sh-NC）的慢病毒感染U251 细胞后，采用实时荧光定量PCR法检测各组细胞中 Mig-7的表达水平；采用体外三维培养和Transwell小室侵袭实验观察Mig-7基因沉默对各组U251细胞VM 形成能力和侵袭能 力的影响。采用western-blot检测各组细胞中MEK/ERK的蛋白表达水平。结果携带有sh-Mig-7和sh-NC的慢病毒成功感染 U251细胞，并获得稳定低表达Mig-7基因的U251细胞株；与感染sh-NC慢病毒和未感染病毒的的细胞相比，sh-Mig-7感染组 U251细胞中Mig-7的表达水平以显著降低（P均<0.01）；与空白对照组和sh-NC感染组相比，sh-Mig-7 感染组U251细胞的侵袭 能力明显下降（P<0.01），并且sh-Mig-7 感染组U251 细胞VM形成能力明显下降（P<0.05）。与空白对照组和sh-NC感染组相 比，sh-Mig-7感染组U251细胞的MEK、ERK蛋白的表达水平均显著降低（P<0.05）。结论沉默Mig -7 基因的表达，可能通过 MEK/ERK信号通路抑制U251 细胞的VM 形成及侵袭能力，提示Mig -7 基因在人脑胶质瘤细胞的VM 和侵袭中发挥重要 的作用。

Mig-7 gene silencing inhibits vasculogenic mimicry formation and invasion of glioma U251 cells in vitro by suppressing MEK/ERK signaling

Objective To investigate the inhibitory effects of silencing migration-inducing gene-7 (Mig-7) on vasculogenic mimicry formation, migration and invasion of human glioma cells in vitro and whether MEK/ERK signaling pathway mediates these effects. Methods Human glioma U251 cells were infected by lentiviral vectors carrying a small interfering RNA targeting Mig-7 gene (sh-Mig-7) or a negative control shRNA (sh-NC), and real-time quantitative PCR was used to detect the expression level of Mig-7 mRNA in the cells. Three-dimensional culture and Transwell chamber invasion assay were used to observe the effect of Mig-7 gene silencing on vasculogenic mimicry formation and invasion ability of the U251 cells. Western blotting was performed to detect the changes in the protein expression levels of MEK/ERK in the infected cells. Results We successfully obtained a U251 cell line with stable low expression of Mig-7 gene using RNA interference technique. Compared with the cells infected with sh-NC lentivirus and the non- infected cells, U251 cells infected with the lentiviral vector carrying sh-Mig-7 showed significantly decreased expression level of Mig-7 (P<0.01) with obviously lowered vasculogenic mimicry formation and invasion abilities (P<0.05). Mig-7 silencing also significantly lowered the expressions of MEK and ERK proteins in U251 cells (P<0.05). Conclusion Silencing of Mig-7 gene inhibits vasculogenic mimicry formation and invasion of U251 cells possibly by suppressing MEK/ERK signaling, suggesting the important role of Mig-7 gene in vasculogenic mimicry formation and invasion of human glioma cells.