摘要
目的了解磷酸肌醇4磷酸(PI4P)对人脑胶质瘤细胞侵袭迁移的影响,探索PI4P在人脑胶质瘤发生过程中的作用机制。方法包装慢病毒,构建U87-GFP(过表达PI4P细胞系对照组)、U87-GFP-PI4P(过表达PI4P细胞系实验组)、U87-Scramble(沉默PI4P细胞系对照组)、U87-sh PI4P(沉默PI4P细胞系实验组)细胞系。应用Werston blot检测各组PI4P的表达水平,应用细胞划痕实验和Transwell实验检测细胞迁移和侵袭能力。结果免疫印迹实验检测出U87-GFP-PI4P细胞系中外源性PI4P表达,而U87-sh PI4P细胞系中PI4P表达较对照组U87-Scramble细胞系减少。过表达PI4P的U87-GFP-PI4P细胞迁移能力较对照组U87-GFP细胞增强(P<0.01);沉默PI4P后迁移能力较对照组减弱(P<0.01)。过表达PI4P的U87细胞侵袭能力强于对照组(P<0.05),沉默PI4P后侵袭能力减弱(P<0.05)。结论 PI4P可促进脑胶质瘤细胞的侵袭迁移,为脑胶质瘤的基础研究及临床治疗提供新的靶点。
Objective To investigate the effect of phosphoinositide 4-phosphate（PI4P） on human glioma U87 cells and the mechanism of action of PI4 P in the development of human glioma through the overexpression or silencing of PI4 P in human glioma U87 cells,and to provide a new target for basic research and clinical treatment of glioma.Methods LV-Helper1,LV-Helper2,p WPXLd-PI4 P,and p LL3.7-sh PI4 P were used to package p WPXLd-PI4 P and p LL3.7-sh PI4 P lentiviruses.The U87-GFP（PI4P-overexpression control group）,U87-GFP-PI4P（PI4P-overexpression experimental group）,U87-Scramble（PI4P-silencing control group）,and U87-sh PI4P（PI4P-silencing experimental group） cell lines were established.Wound-healing assay and Transwell assay were used to evaluate cell migration and invasion,and Western blot was used to measure the expression of PI4 P in each group.Results Western blot detected the expression of exogenous PI4 P in the U87-GFP-PI4 P cell line,and the U87-sh PI4 P cell line showed reduced expression of PI4 P compared with the U87-Scramble cell line in the control group.The U87-GFP-PI4 P cell line with PI4 P overexpression had a significantly stronger ability of migration than the U87-GFP cell line in the control group（P〈0.01）;the U87-sh PI4 P cell line with PI4 P silencing had a reduced ability of migration than the U87-Scramble cell line in the control group（P〈0.01）.The U87 cell line with PI4 P overexpression had a significantly stronger invasion ability than the control group（P〈0.05）;after PI4 P silencing,the experimental group showed a significant reduction in invasion ability compared with the control group（P〈0.05）.Conclusions In human glioma U87 cells,PI4 P can promote the invasion and migration of glioma cells and may become a new target in the basic research and clinical treatment of glioma.
出处
《中国当代儿科杂志》
CAS
CSCD
北大核心
2016年第8期775-780,共6页
Chinese Journal of Contemporary Pediatrics
