亚硝酸钠诱导人肝癌SMMC-7721细胞上皮-间质转化  

Sodium nitrite induces epithelial-mesenchymal transition of SMMC-7721 cells

作　　者：王予东[1] 付建民[1] 史齐[1] 李延红[1] 皇甫超申[1,2] 

机构地区：[1]河南大学医学院环境医学研究所,河南开封475004 [2]河南大学环境与健康工程中心,河南开封475004

出　　处：《药学学报》2011年第5期507-512,共6页Acta Pharmaceutica Sinica

基　　金：国家重大环保公益项目专项(200809115);省部共建河南大学科研项目(SBGJ090702)

摘　　要：探讨亚硝酸盐诱导人肝癌SMMC-7721细胞发生上皮-间质转化(EMT)的作用。0.25～25 mmol·L-1亚硝酸钠处理细胞48 h后,酶联免疫吸附实验(ELISA)检测细胞培养上清液中的TGF-β1、IL-6和IL-8水平,相差倒置显微镜下观察细胞形态学变化,划痕实验和Transwell侵袭实验检测细胞移动和侵袭能力,同时采用Western blotting方法检测EMT相关标记蛋白和p-AKT及下游信号分子的蛋白表达水平。结果表明,亚硝酸钠处理人肝癌细胞48 h,可以引起TGF-β1的分泌显著升高,并呈现剂量依赖性。亚硝酸钠对IL-8及IL-6的分泌也具有一定的增加作用,但不具有剂量依赖性。0.25 mmol·L-1亚硝酸钠作用48 h,诱导细胞形态向间质转化,增强波形蛋白和p-AKT及其下游信号蛋白的表达,抑制E型钙黏素蛋白的表达,促进细胞迁移和侵袭能力。结果提示,亚硝酸钠可能通过促进细胞分泌TGF-β1和IL-8,诱导人肝癌SMMC-7721细胞上皮-间质转化。This study is to find out the induction by sodium nitrite of epithelial-mesenchymal transition （EMT） in human hepatocellular carcinoma cells, SMMC-7721. After treatment of SMMC-7721 with 0.25-25 mmol.L^-1 sodium nitrite for 48h, the assays used include enzyme-linked immunosorbent assay （ELISA） for evaluation of TGF-βl, IL-6 and IL-8 level in the conditioned medium, phase-contrast microscopy for morphology observation, and scratch wound healing as well as transwell migration assays for measurement of migration and metastatic potential. Additionally, the hallmarks of EMT, p-AKT and its downstream signaling molecules were examined by Western blotting. The results showed that TGF-β1 secreted by SMMC-7721 elevated significantly in a dose-dependent fashion, whereas the increased IL-8 and IL-6 did not show dose-dependent response. The EMT was induced by exposure of SMMC-7721 with 0.25 mmol＇L^-l of sodium nitrite, which was characterized by increased level of Vimentin, decreased E-cadherin and elevated activity of migration and metastatic potential. The results suggest that sodium nitrite could induce SMMC-7721 EMT by increased secretion of TGF-β1 and IL-8

关 键 词：亚硝酸钠 人肝癌细胞 细胞因子 上皮-间质转化 

分 类 号：R963[医药卫生—微生物与生化药学]