基础医学与临床 ›› 2017, Vol. 37 ›› Issue (1): 19-24.

• 研究论文 • 上一篇    下一篇

血管细胞黏附分子-1增强人脑胶质瘤细胞系细胞迁移侵袭能力

钟杰1,曲彦明2,韩松3,罗艳琳1,李俊发1   

  1. 1. 首都医科大学
    2. 首都医科大学第十一临床医学院,北京三博脑科医院
    3. 首都医科大学神经生物学系
  • 收稿日期:2016-07-29 修回日期:2016-09-26 出版日期:2017-01-05 发布日期:2016-12-30
  • 通讯作者: 李俊发 E-mail:junfali@ccmu.edu.cn
  • 基金资助:
    国家自然科学基金面上项目;北京市自然科学基金资助项目

VCAM-1 enhances the migration activity and invasion ability of human glioma cell line cells

  • Received:2016-07-29 Revised:2016-09-26 Online:2017-01-05 Published:2016-12-30
  • Contact: Jun-fa LI E-mail:junfali@ccmu.edu.cn

摘要: 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T98G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。

关键词: 血管细胞黏附分子-1, 脑胶质瘤, 慢病毒转染, 迁移能力, 侵袭能力

Abstract: 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 EF1a-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 the migration activity and invasion ability 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 T98G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were firstly 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 enhances the migration activity and invasion ability of human glioma cell line cells.

Key words: Vascular cell adhesion molecule-1 (VCAM-1), brain glioma, lentiviral transfection, migration activity, invasion ability.

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