학술논문
血管细胞黏附分子-1增强人脑胶质瘤细胞系迁移和侵袭能力 / VCAM-1 improves migration and invasion of human glioma cell lines
Document Type
Academic Journal
Author
Source
基础医学与临床. 37(1):19-24
Subject
Language
Chinese
ISSN
1001-6325
Abstract
目的：探讨血管细胞黏附分子-1（VCAM-1）在脑胶质瘤细胞迁移侵袭能力中作用。方法用慢病毒pSGU6／GFP／Neo介导VCAM-1的shRNA、慢病毒EF1a-GFP／puro介导VCAM-1过表达载体、划痕迁移、Transwell 侵袭、蛋白免疫印迹（Western blot）和细胞染色等实验技术和方法，观察了VCAM-1蛋白表达水平对人脑胶质瘤T98G和U251细胞系细胞迁移和侵袭能力的影响。其中，T98G细胞分为空白对照组、空载体对照组、乱序对照组和实验组（抑制VCAM-1蛋白表达水平组），U251细胞分为空白对照组、空载体对照组和实验组（过表达VCAM-1组），每组6个复孔。结果首先利用慢病毒介导VCAM-1的shRNA和过表达载体建立了稳定低表达VCAM-1的T98G细胞和稳定过表达VCAM-1的U251细胞。稳定低表达VCAM-1的T98 G细胞划痕恢复能力（迁移能力）明显减弱（ P＜0.01）；而稳定过表达VCAM-1的U251细胞迁移能力明显提高（P＜0.05）。同样，稳定低表达VCAM-1的T98G细胞侵袭能力显著减弱（ P ＜0.05）；而稳定过表达 VCAM-1的 U251细胞侵袭能力明显增强（ P ＜0.01）。结论VCAM-1可显著增强人脑胶质瘤细胞系细胞的迁移和侵袭能力。
Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.
Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.