sunitinib对气道平滑肌细胞增殖和迁移的影响及机制

The effect and mechanisms of sunitinib on airway smooth cells proliferation and migration

目的:探讨sunitinib(sunitinib malate)对血小板源性生长因子BB(PDGF-BB)刺激的人气道平滑肌细胞(human airway smooth muscle cell,HASMCs)增殖和迁移的影响及其机制。方法:体外培养HASMCs分为:对照组、PDGF-BB组、PDGF-BB与sunitinib干预组、sunitinib组,四甲基偶氮唑蓝(MTT)微量比色法测定HASMCs增殖,流式细胞术检测HASMCs细胞周期,改良的Boyden微孔膜双槽法观察细胞迁移,Westernblot检测PDGFR-β和AKT的磷酸化。结果:与对照组相比,PDGF-BB(20ng/ml)显著诱导HASMCs的增殖和迁移(P<0.05),sunitinib(0.3～9.0nmol/L)呈浓度依赖性抑制PDGF-BB诱导的HASMCs增殖和迁移;PDGF-BB(20ng/ml)刺激HASMCs后,细胞周期S期比例显著高于对照组(P<0.05),PDGF-BB与sunitinib干预组细胞周期S期比例低于PDGF-BB组(P<0.05);PDGF-BB组PDGFR-β和AKT的磷酸化水平较对照组为高(P<0.05),PDGF-BB与sunitinib干预组其表达量低于PDGF-BB组。结论:sunitinib抑制PDGF-BB诱导的HASMCs增殖和迁移,可能是通过调节PI3K/AKT通路起作用。

Objective：To investigate the effect and mechanisms of sunitinib on PDGF-BB-induced proliferation and migration of human airway smooth muscle cells（HASMCs）.Methods：HASMCs derived from human bronchus were divided into four groups：normal group,PDGF-BB group,PDGF-BB and sunitinib intervention group,sunitinib group.Proliferation was measured by MTT.Cell cycle progression was analyzed by flow cytometry.Migration was examined using Boyden chamber apparatus.The protein expressions of p-PDGFR-βand p-AKT were determined by western blot analysis.Results：PDGF-BB（20 ng /ml） significantly induced proliferation and migration of HASMCs as compared to controls.Sunitinib（0.3～9 nmol /L） inhibited the HASMCs proliferation and migration in response to PDGF-BB（20 ng /ml）.Flow cytometry disclosed that PDGF-BB（20 ng /ml） increased significantly HASMCs ratio in S phase（P 〈0.05）;Sunitinib decreased HASMCs in S phase stimulated by PDGF-BB（P〈 0.05）.The phosphorylated protein levels of PDGFR-β and AKT in PDGF-BB group were higher than those of the normal group（P 〈0.05）,but were decreased in PDGF-BB and sunitinib intervention group compared with control（P〈 0.05）.Conclusion：It appears that sunitinib can directly inhibit PDGF-BB induced HASMCs proliferation and migration through the modulation of the PI3K /Akt pathway.