摘要
目的研究高良姜素对胶质瘤细胞迁移和侵袭的影响并初步探讨其作用机制。方法（1）体外常规培养人脑胶质瘤细胞系U87、U251，CCK．8法检测0、10、20、40μmol／L高良姜素作用24h后细胞存活率的变化：伤口愈合实验、Transwell实验、Western blotting实验分别检测0、10、20μmol／L高良姜素作用24h后细胞迁移、侵袭能力的变化和转化生长因子β（TGFβ）的表达；（2）实验分为对照组、20jxm01／L高良姜素组、TGF[3过表达组、TGFB过表达＋20μmol／L高良姜素组。前2组细胞转染空载质粒、后2组细胞转染TGFβ质粒，转染后24h第2、4组细胞均加入20μmol／L高良姜素．继续培养24h后采用伤口愈合实验、Transwell实验、Western blotting实验分别检测4组细胞的迁移、侵袭能力和TGFβ的表达。结果（1）与0、10、20μmol／L高良姜素组比较，40μmol／L高良姜素组细胞存活率降低，差异有统计学意义（P〈0．05）。0、10、20μmol／L高良姜素组U87、U251细胞的伤口愈合百分比均依次下降，侵袭到达下室的细胞逐渐减少，TGFβ蛋白表达量逐渐下降，差异有统计学意义（P〈0．05）；（2）TGFβ过表达组、对照组、TGFβ过表达＋20μmol／L高良姜素组、20μmol／L高良姜素组U87、U251细胞的伤口愈合百分比依次下降，侵袭到达下室的细胞逐渐减少，TGFβ蛋白表达量逐渐下降（U87细胞TGFβ蛋白表达量分别为1．63±0．21、1．00±0．00、0．78±0．05、0．43＋0．08；U251细胞TGFβ蛋白表达量分别为1．98±0．20、1．00±0．00、0．86±0．06、0．29±0．04），差异均有统计学意义（P〈0．05）。结论高良姜素可以通过阻断TGFβ来抑制胶质瘤母细胞U87和U251的迁移和侵袭。
Objective To investigate the effect of Galangin on migration and invasion in gliomas and explore the possible mechanism. Methods （1） After treating the glioma lines U87 and U251 with 0, 10, 20 and 40 μmol/L galangin for 24 h, CCK-8 was performed to detect the cell viability; wound healing assay and Transwell assay were used to detect the cell migration and invasion, respectively; protein expression of transforming growth factor （TGF） β was detected by Western blotting. （2） The second experiment was divided into control group, 20 μmol/L galangin treatment group, TGFβ over-expression group and 20 μmol/L galangin±TGFβ over-expression group; the cells in the control group and 20 μmol/L galangin treatment group were transfected with empty-vector plasmids; cells in the TGFβ over-expression group and 20 μmol/L galangin±TGFβ over-expression group were transfected with TGFβ plasmids, and 24 h after the transfection, the cells in the 20 μmol/L galangin treatment group and 20 μmol/L galangin＋TGFβ over-expression group were added 20 μmol/L galangin; 24 h after that, wound healing assay and Transwell assay were used to detect the cell migration and invasion,respectively; TGF β protein expression was detected by Western blotting. Results （1） As compared with cells from the 0, 10 and 20 μmol/L galangin group, the cells from 40 μmol/L galangin group had significantly lower cell viability （P〈0.05）. Cells from the 0, 10 and 20 μmol/L galangin group had significantly decreased wound healing percentage, significantly decreased cell number reaching to the lower chamber, and statistically decreased TGFI3 protein expression in sequence （P〈0.05）. （2） U87 and U251 cells from the TGFβ over-expression group, control group, 20 μmol/L galangin±TGFβ over-expression group and 20 μmol/L galangin treatment group had successively decreased wound healing percentage, decreased cell number reaching to the lower chamber, and decreased TGFβ protein expression （U87 cells： 1.63±0.21, 1.00±0.00, 0.78±0.05 and 0.43±0.08; U251 cells： 1.98±0.20, 1.00± 0.00, 0.86±0.06 and 0.29±0.04）, with significant differences （P〈0.05）. Conclusion Galangin inhibits the glioma migration and invasion through TGFbβ down-regulation.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2016年第9期871-877,共7页
Chinese Journal of Neuromedicine
基金
四川省科学技术厅重点科技自筹项目（2013SZZ002）
西南医科大学青年基金项目（2013ZRQN068）
四川省科学技术厅与泸州市人民政府、西南医科大学联合科研专项资金计划项目（20147SX-0030）
关键词
高良姜素
迁移
侵袭
转化生长因子Β
Galangin
Migration
Invasion
Transforming growth factor β
