摘要
探讨盐霉素对人肝癌细胞HepG2增殖、侵袭转移能力影响的作用机制。MTT法检测盐霉素对肝癌HepG2细胞和正常肝细胞L02增殖的影响;利用FITC标记的鬼笔环肽进行微丝免疫荧光染色,激光共聚焦技术获得盐霉素作用下F-actin细胞骨架的形态变化;Transwell小室法测定盐霉素对HepG2细胞体外迁移侵袭能力的变化。Western blot法检测盐霉素对HepG2细胞中β-连环蛋白(β-catenin)、基质金属蛋白酶-2(MMP-2)及基质金属蛋白酶-9(MMP-9)蛋白表达的影响。结果显示,盐霉素抑制HepG2细胞的增殖,并呈现出时间、剂量依赖效应;而盐霉素对正常肝细胞L02的增殖抑制作用不明显。经过盐霉素(1μmol/L和4μmol/L)处理的HepG2细胞,迁移和侵袭实验中穿膜细胞数均明显低于对照组(P<0.05),且以F-actin为基础的微丝骨架结构发生紊乱。盐霉素(4μmol/L和8μmol/L)还可明显下调MMP-2、MMP-9及β-catenin蛋白的表达水平(P<0.05)。这些结果提示,盐霉素能有效抑制人肝癌细胞HepG2的增殖、迁移和侵袭,其作用机制可能与降低MMP-2、MMP-9及β-catenin蛋白的表达有关。
To investigate effects of salinomycin on the proliferative, invasive and metastatic abilities of hepatoma cell line HepG2 in vitro and analyze the possible mechanisms, MTT was used for determining the prolif- erative abilities of HepG2 and human normal liver L02 ceils. The cytoskeleton was observed using confocal laser scanning microscopy after F-actin staining by FITC-labeled phalloidin. The invasive and migratory abilities were detected by Transwell assay. The protein expressions of 13-catenin, MMP-2 and MMP-9 at protein levels were eval- uated by Western blot. The results showed that the treatment of L02 cells with salinomycin （0-8 ~tmol/L） for 24 h and 48 h had no significant effects on cell viability. However, treatment of similar doses of salinomycin suppressedthe growth of HepG2 cells in a time and concentration dependent manner. HepG2 was treated by salinomycin （I, 4 ~tmol/L） for 24 h before invasive and migratory abilities were examined. Compared with that in the control group, the number of invasive and migratory cells was remarkably decreased （P〈0.05）, the structure of F-actin micro filament cytoskeleton was significantly changed. In addition, immuno-blot showed that the expressions of 13-catenin, MMP-2 and MMP-9 at protein levels were significantly down-regulated （P〈0.05）. The results suggested that the proliferative, invasive and migratory abilities of hepatoma cells could be inhibited by salinomycin, which may be related to down-regulating the protein expressions of 13-catenin, MMP-2 and MMP-9.
出处
《中国细胞生物学学报》
CAS
CSCD
北大核心
2013年第5期655-660,共6页
Chinese Journal of Cell Biology
基金
兰州大学中央高校基本科研业务费专项资金(批准号:lzujbky-2010-127)资助的课题~~
关键词
盐霉素
肝癌细胞
增殖
侵袭转移
体外实验
salinomycin
hepatoma cell
proliferation
invasion and metastasis
in vitro experiment
