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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。
To investigate the role of nitrite-induced epithelial-mesenchymal transition (EMT) in human hepatoma SMMC-7721 cells. The cells were treated with 0.25-25 mmol·L-1 sodium nitrite for 48 h, and the levels of TGF-β1, IL-6 and IL-8 in the cell culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA) Morphological changes, scratch test and Transwell invasion assay were used to detect cell migration and invasion. Meanwhile, Western blotting was used to detect the expression of EMT-related marker proteins and p-AKT and downstream signaling molecules. The results showed that sodium nitrite treatment of human hepatocellular carcinoma cells 48 h, can cause TGF-β1 secretion was significantly increased, and showed a dose-dependent manner. Sodium nitrite also increased the secretion of IL-8 and IL-6, but not in a dose-dependent manner. 0.25 mmol·L-1 sodium nitrite for 48 h, inducing the transformation of cell morphology into stroma, enhancing the expression of vimentin and p-AKT and its downstream signaling proteins, inhibiting the expression of E-cadherin and promoting cell migration and Invasive ability. The results suggest that sodium nitrite may induce epithelial-mesenchymal transition in human hepatoma SMMC-7721 cells by promoting the secretion of TGF-β1 and IL-8 by cells.