钾通道阻断剂对胶质瘤细胞迁移和侵袭的影响

Effect of Potassium Channel Blockers on Migration and Invasion of Human Glioma Cells

为了检测电压门控钾通道阻断剂4-氨基吡啶(4-AP)、四乙胺(TEA)和ATP敏感钾通道阻断剂格列苯脲(Glibenclamide,Gli)对胶质瘤细胞迁移和侵袭的影响,选用人胶质瘤细胞系U87和U251,其中钾通道阻断剂4-AP、TEA及Gli处理作为实验组,未处理的作为对照组.采用划痕实验和Transwell小室法检测钾通道阻断剂对U87和U251细胞迁移和侵袭能力的影响;Western blot检测药物处理后细胞高迁移率蛋白B1(high mobility group protein B1,HMGB1)表达水平.结果表明:5 mmol·L^(-1)的4-AP、40 mmol·L^(-1)的TEA及400μmol·L^(-1)的Gli可以显著抑制胶质瘤细胞的迁移、侵袭,并降低HMGB1表达水平.电压门控钾通道和ATP敏感钾通道对胶质瘤细胞迁移和侵袭具有重要调控作用,3种钾通道阻断剂对胶质瘤细胞迁移和侵袭有不同程度的抑制作用,可能通过调控HMGB1相关通路实现.

To explore the effect of potassium channel blocker on glioma migration and invasion,voltage-gated potassium channel blocker 4-aminopyridine,tetraethylammonium and ATP sensitive potassium channel blocker glibenclamide were investigated.Human glioma cell lines U87 and U251 were selected to use.Wound-healing assay and Transwell assays were performed to determine the migration and invasion of glioma cells.The expression level of HMGB1 protein was detected by western blot.Cell abilities of migration and invasion were significantly reduced with4-AP（5 mmol·L-1）,TEA（40 mmol·L-1） or glibenclamide（400 μmol·L-1） compared with that of the control.The expression level of HMGB1 was significantly down-regulated in the blockers groups.Potassium channels play important roles in the migration and invasion of human glioma.The selective blockers of voltage-gated potassium channels or ATP-sensitive potassium channels significantly inhibited the migration and invasion of U87 and U251 cells,and down-regulation of HMGB1 gene expressions might be its possible mechanism.