摘要
目的初步研究PPTA对肝癌细胞HCCLM3增殖、周期和迁移的作用及机制.方法分别采用四甲基偶氦唑蓝法(methyl thiazolyl tetrazolium,MTT)、流式细胞术、细胞划痕实验,观察PPTA对肝癌细胞HCCLM3增殖、周期和迁移的影响;采用细胞免疫荧光实验法观察PPTA对HCCLM3细胞肌动蛋白纤维的分布、丝状和片状伪足形成等的影响;采用Western blot法检测肝癌细胞HCCLM3中蛋白激酶B(protein kinase B,AKT)和细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)的表达.结果PPTA对HCCLM3细胞有抑制作用,抑制HCCLM3细胞增殖的IC_(50)在50μmol/L以下.当PPTA浓度为40μmol/L时,对HCCLM3细胞周期有抑制作用.PPTA对HCCLM3细胞划痕的相对愈合速度有抑制作用,随着浓度的升高抑制作用显著增强.细胞免疫荧光显示,HCCLM3细胞中丝状伪足增多,片状伪足减少,细胞排列更为紧密,当PPTA浓度为40μmol/L,作用显著;Western blot显示,当PPTA浓度为20、40μmol/L时,上调HCCLM3细胞的AKT表达,下调pAKT表达.结论PPTA抑制肝癌细胞HCCLM3的增殖、周期、迁移,其作用机制可能是调控AKT和ERK相关的信号通路.
AIM To investigate the effects of pholidotaphenanthrene A（PPTA） on the proliferation,apoptosis,migration,and cell cycle progression of hepatocellular carcinoma HCCLM3 cells and the underlying mechanism.METHODS Methyl thiazolyl tetrazolium（MTT） assay,flow cytometry and wound healing assay were used to evaluate the effects of PPTA on the viability,cell cycle progression and migration of HCCLM3 cells,respectively.Immunofluorescence staining and Western blot assays were performed to explore the effects of PPTA on F-actin distribution,the formation of lamellipodia and filopodia,and the expression of related signaling molecules.RESULTS PPTA decreased the viability of HCCLM3 cells,with IC（50） 50 μmol/L.PPTA at a dose of 40 μmol/L obviously delayed the cell cycle progression of HCCLM3 cells.Results of wound healing assay showed that PPTA can inhibit the relative healing rate of HCCLM3 cells in a dose-dependent manner.Immunofluorescence staining showed that HCCLM3 cells after incubation with different concentrations of PPTA displayed more close arrangement,decreased lamellipodia formation and increased filopodia formation,especially in cells treated with PPTA at a dose of 40 μmol/L.Western blot analysis showed that PPTA at 20 and40 μmol/L can up-regulate protein kinase B（AKT） and down-regulate pAKT expression.CONCLUSION PPTA can inhibit the proliferation,cycle progression,and migration HCCLM3 cells possibly via mechanisms related to altering the levels of pAKT and pERK.
出处
《世界华人消化杂志》
CAS
2016年第30期4177-4184,共8页
World Chinese Journal of Digestology
关键词
PPTA
肝癌
HCCLM3细胞
增殖
周期
迁移
Pholidotaphenanthrene A
Hepatocellular carcinoma cells
HCCLM3 cells
Proliferation
Cell cycle
Migration
