As_2O_3联合AZT作用对肝癌HepG2细胞迁移和侵袭的影响  

Effects of As_2O_3 combined with AZT on migration and invasion of liver cancer HepG2 cells

作　　者：韩丽[1] 陈彻[1] 楚惠媛[1] 刘玉[1] 梁永娟[1] 李晶莹[1] 

机构地区：[1]甘肃中医药大学医学技术学院,甘肃兰州730000

出　　处：《肿瘤》2015年第9期982-989,共8页Tumor

基　　金：国家自然科学基金资助项目(编号:81460456);甘肃省自然科学基金项目(编号:1308RJZA169)~~

摘　　要：目的 :观察三氧化二砷(arsenic trioxide,As2O3)联合3’-叠氮-3’-脱氧胸腺嘧啶核苷(3’-azido-3’-deoxythymidine,AZT)对人肝癌Hep G2细胞迁移和侵袭的影响,并探讨其可能的作用机制。方法 :分别用As2O3、AZT以及两药联合处理肝癌Hep G2细胞,同时以未经药物处理的Hep G2细胞作为空白对照。应用划痕愈合实验和Transwell迁移及侵袭实验分别检测细胞迁移及侵袭能力的变化,实时荧光定量PCR法检测细胞中基质金属蛋白酶2(matrix metallopeptidase 2,MMP2)和血管内皮生长因子(vascular endothelial growth factor,VEGF)m RNA表达的变化,蛋白质印迹法检测MMP2、VEGF、细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK1/2)和磷酸化ERK1/2(phosphorylated ERK1/2,p-ERK1/2)蛋白表达的变化。结果 :经As2O3联合AZT干预后,Hep G2细胞的迁移和侵袭能力均比空白对照及单药组明显降低(P值均<0.01)。As2O3联合AZT干预组Hep G2细胞中,MMP2和VEGF m RNA的表达水平明显低于空白对照组和各单药组(P值均<0.01),MMP2、VEGF和p-ERK1/2蛋白的表达水平也明显降低(P值均<0.01),而ERK1/2蛋白表达无明显变化(P>0.05)。结论 :As2O3联合AZT作用能够明显抑制人肝癌Hep G2细胞的迁移和侵袭能力,该抑制作用可能是通过调控ERK1/2信号通路的磷酸化以及下调MMP2和VEGF的表达来实现的。Objective： To investigate the effects of arsenic trioxide （As2O3） combined with 3＇-azido-3＇-deoxythymidine （AZT） on the migration and invasion of human liver cancer HepG2 cells, and to explore its possible mechanism.Methods： The liver cancer HepG2 cells were treated with AszO3 or AZT alone or in combination; meanwhile, the HepG2 cells without any treatment were used as the blank control. The migration and invasion of HepG2 cells were measured by wound healing assay and Transwell migration and invasion assays, respectively. The expression mRNA levels of matrix metallopeptidase 2 （MMP2） and vascular endothelial growth factor （VEGF） were detected by real-time fluorescent quantitative-PCR. The protein expression levels of MMP2, VEGF, extracellular signal-regulated kinase 1/2 （ERK1/2） and phosphorylated ERK1/2 （p-ERK1/2） were detected by Western blotting.Results： The migration and invasion abilities of HepG2 cells in As203 combined with AZT group were significantly decreased than those in the blank control, As2O3 and AZT alone groups （all P 〈 0.01）. As compared with the blank control, As2O3 and AZT alone groups, the mRNA expression levels of VEGF and MMP2 in As203 combined with AZT group were down- regulated （all P 〈 0.01）, as well as the protein expression levels of MMP2, VEGF and p-ERK1/2 were also down-regulated （all P 〈 0.01）; but there was no obvious change in expression of ERK1/2 protein among four groups （P 〉 0.05）.Conclusion： As2O3 combined with AZT has synergistic inhibitory effects on the migration and invasion abilities of HepG2 cells. The effect may be related to phosphorylation of ERK1/2 pathway and down-regulation of MMP2 and VEGF expressions.

关 键 词：肝肿瘤 药物筛选试验 抗肿瘤 肿瘤浸润 基因表达调控 肿瘤 三氧化二砷 3’-叠氮-3’-脱氧胸腺嘧啶核苷 

分 类 号：R735.7[医药卫生—肿瘤]