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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 the effect of salinomycin on the proliferation, invasion and metastasis of HepG2 human hepatocellular carcinoma cells. MTT assay was used to detect the effect of salinomycin on the proliferation of HepG2 cells and normal liver cells L02. FITC-labeled phalloidin was used to immunofluorescence staining and confocal laser scanning microscopy to obtain the cytoskeleton of F-actin Morphological changes; Transwell chamber method to determine the salinomycin on HepG2 cells in vitro migration and invasion changes. The effect of salinomycin on the protein expression of β-catenin, matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in HepG2 cells was detected by Western blot. The results showed that salinomycin inhibited the proliferation of HepG2 cells in a time-and dose-dependent manner, whereas salinomycin had no obvious inhibitory effect on the proliferation of L02 cells. In HepG2 cells treated with salinomycin (1μmol / L and 4μmol / L), the number of transmembrane cells in migration and invasion assays was significantly lower than that in the control group (P <0.05), and F-actin- Structural disorder. Salinomycin (4μmol / L and 8μmol / L) also significantly downregulated the expression of MMP-2, MMP-9 and β-catenin (P <0.05). These results suggest that salinomycin can effectively inhibit the proliferation, migration and invasion of HepG2 cells, which may be related to the decrease of the expression of MMP-2, MMP-9 and β-catenin.