磷酸肌醇4磷酸对人脑胶质瘤U87细胞侵袭迁移的影响

Effect of phosphoinositide 4-phosphate on invasion and migration of human glioma U87 cells

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-shPI4P （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-shPI4P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。

Abstract:

Objective To investigate the effect of phosphoinositide 4-phosphate (PI4P) on human glioma U87 cells and the mechanism of action of PI4P in the development of human glioma through the overexpression or silencing of PI4P in human glioma U87 cells, and to provide a new target for basic research and clinical treatment of glioma. Methods LV-Helper1, LV-Helper2, pWPXLd-PI4P, and pLL3.7-shPI4P were used to package pWPXLd-PI4P and pLL3.7-shPI4P lentiviruses. The U87-GFP (PI4P-overexpression control group), U87-GFP-PI4P (PI4P-overexpression experimental group), U87-Scramble (PI4P-silencing control group), and U87-shPI4P (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 PI4P in each group. Results Western blot detected the expression of exogenous PI4P in the U87-GFP-PI4P cell line, and the U87-shPI4P cell line showed reduced expression of PI4P compared with the U87-Scramble cell line in the control group. The U87-GFP-PI4P cell line with PI4P overexpression had a significantly stronger ability of migration than the U87-GFP cell line in the control group (P < 0.01); the U87-shPI4P cell line with PI4P 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 PI4P overexpression had a significantly stronger invasion ability than the control group (P < 0.05); after PI4P 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, PI4P can promote the invasion and migration of glioma cells and may become a new target in the basic research and clinical treatment of glioma.