目的探讨溶血磷脂酸(LPA)诱导HeLa-S3细胞迁移能力的机制。方法采用细胞迁移实验测定HeLa-S3细胞的迁移;Western blot检测Akt、P-Akt(Ser473)、PAK1、P-PAK1(Thr423)的表达;采用CM-H2DCFDA检测细胞ROS的生成情况。结果①20μmol·L-1LPA可使细胞内ROS的产生明显增加(P〈0.05),采用NAC预处理后能明显的抑制ROS的生成;②LPA可升高P-Akt(Ser473),P-PAK1(Thr423)/PAK1水平,但采用LY294002可以抑制PAK1(Thr423)磷酸化水平的升高和细胞的迁移(P〈0.01)。结论溶血磷脂酸通过上调ROS水平来诱导HeLa-S3细胞的迁移,其可能的机制是通过PI3K信号通路活化Akt/PAK1来实现的。
Aim To explore the mechanisms of lysophosphatidic acid(LPA) in the migration of HeLa-S3 cell.Methods The cell migration experiments were employed to test the migration of HeLa-S3 cells;Western blot was used to analyse Akt,P-Akt(Ser473),PAK1,P-PAK1(Thr423) expression;CM-H2DCFDA was used to test the Intracellular ROS level.Results(1) 20 μmol·L-1 LPA increased the production of intracellular ROS(P0.05),while the pretreatment with NAC significantly inhibited LPA-induced intracellular ROS generation;(2) LPA elevated-Akt(Ser473)/Akt and P-PAK1(Thr423)/PAK1,while the pretreatment with LY294002 inhibited the increased P-PAK1(Thr423)/PAK1and cell migration.Conclusions LPA induces the migration of HeLa-S3 cells through the up-modulation of ROS level,and its possible mechanisms may be related to activating Akt/PAK1 by PI3K signal pathway.