喹唑啉类PAK4小分子抑制剂影响胰腺癌细胞迁移及侵袭的作用机制

The mechanism of quinazoline PAK4 small molecule inhibitor affecting the migration and invasion of pancreatic cancer cells

目的:探究喹唑啉类PAK4小分子抑制剂Czh#226对胰腺癌细胞增殖、迁移等方面的作用。方法:四唑盐比色法(MTT)检测不同浓度的Czh#226对胰腺癌细胞增殖的影响;Transwell实验检测不同浓度Czh#226对胰腺癌细胞迁移的影响;激光扫描共聚焦显微(Confocal)实验检测不同浓度Czh#226对胰腺癌细胞微丝的影响;Western blot实验检测不同浓度Czh#226对胰腺癌细胞PAK4激酶的活化及其下游迁移侵袭相关蛋白的表达。结果:Czh#226对胰腺癌细胞的增殖无抑制作用,但在5μmol/L时对迁移及侵袭能力有明显抑制作用;Czh#226对PAK4激酶的活性以及PAK4下游细胞骨架相关靶蛋白的磷酸化亦有抑制作用,且均呈剂量效应关系。结论:Czh#226是通过抑制PAK4激酶的活性以及PAK4下游细胞骨架相关靶蛋白的表达来抑制微丝的重组,从而抑制胰腺癌细胞的迁移及侵袭能力。

Objective:To investigate the effects of the quinazoline PAK4 small molecule inhibitor Czh#226 on the proliferation and migration of pancreatic cancer cells lines.Methods:MTT tetrazolium salt colorimetry was used to detect the effects of different concentrations of Czh#226 on the proliferation of pancreatic cancer cells.The effect of different concentrations of Czh#226 on the migration of pancreatic cancer cells was detected by Transwell assay.Confocal assay was used to detect different concentrations of Czh#226 on pancreatic cancer cells ’microfilaments.Western blot assay was used to detect the expression of the activation of PAK4 and the proteins related with migration and invasion in pancreatic cancer cells by different concentrations of Czh#226.Results:Czh#226 had no inhibitory effect on the proliferation of pancreatic cancer cells,but it could inhibit the migration and invasion of pancreatic cancer cells at 5 μmol/L.It also inhibited the expression of the activity of PAK4 kinase and the downstream of PAK4 proteins related with cytoskeleton,and all of them showed a dose-effect relationship.Conclusion:Czh#226 inhibits the reorganization of microfilaments by inhibiting the expression of the activity of PAK4 kinase and the downstream proteins of PAK4 which related to cytoskeleton,thereby inhibiting the migration and invasion of pancreatic cancer cells.