王予东 付建民 史 齐 李延红 皇甫超申. 亚硝酸钠诱导人肝癌SMMC-7721细胞上皮-间质转化J. 药学学报, 2011,46(5): 507-512.
引用本文: 王予东 付建民 史 齐 李延红 皇甫超申. 亚硝酸钠诱导人肝癌SMMC-7721细胞上皮-间质转化J. 药学学报, 2011,46(5): 507-512.
WANG Yu-Dong, Fu-Jian-Min, Shi- Ji, Li-Yan-Gong, Huang-Fu-Chao-Shen. Sodium nitrite induces epithelial-mesenchymal transition of SMMC-7721 cellsJ. 药学学报, 2011,46(5): 507-512.
Citation: WANG Yu-Dong, Fu-Jian-Min, Shi- Ji, Li-Yan-Gong, Huang-Fu-Chao-Shen. Sodium nitrite induces epithelial-mesenchymal transition of SMMC-7721 cellsJ. 药学学报, 2011,46(5): 507-512.

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

  • 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。0.2525 mmol·L−1亚硝酸钠处理细胞48 h, 酶联免疫吸附实验 (ELISA) 检测细胞培养上清液中的TGF-β1IL-6IL-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-8IL-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-β1IL-8, 诱导人肝癌SMMC-7721细胞上皮-间质转化。

     

    Abstract:

    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 48 h, the assays used include enzyme-linked immunosorbent assay (ELISA) for evaluation of TGF-β1, 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−1 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.

     

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