论文部分内容阅读
目的研究酸性环境对肝癌HepG2细胞上皮细胞间质转化(EMT)的影响。方法分别在酸性和碱性条件下培养HepG2细胞,观察细胞形态的区别。划痕实验检测两组细胞的迁移能力,Matrigel侵袭实验检测细胞的侵袭能力。RT-PCR检测两组细胞EMT相关基因mRNA的表达水平,Western blot检测两组细胞EMT相关基因蛋白的表达水平。结果酸性条件下培养的HepG2细胞形态明显从上皮型转变为间质型;酸性条件下HepG2细胞的迁移能力明显强于碱性条件下;酸性条件下HepG2细胞穿越Matrigel的数量明显多于碱性条件下;酸性条件下HepG2细胞EMT相关基因Vimentin、Slug、Snail及Zeb1的mRNA表达以及EMT相关蛋白Vimentin和MMP9的表达强度均明显高于碱性条件下。结论酸性环境能诱导肝癌HepG2细胞EMT的发生。
Objective To study the effect of acidic environment on the epithelial cell mesenchymal transition (EMT) in HepG2 hepatocellular carcinoma. Methods HepG2 cells were cultured under acidic and alkaline conditions respectively to observe the difference of cell morphology. Scratch assay was used to detect the migration ability of the two groups of cells, and Matrigel invasion assay was used to detect the invasion ability of the cells. RT-PCR was used to detect the expression of EMT-related gene mRNA in both groups. Western blot was used to detect the expression of EMT-related genes in both groups. Results The morphology of HepG2 cells cultured in acidic condition evidently changed from epithelial to mesenchymal type. The migration ability of HepG2 cells under acid condition was obviously stronger than that under alkaline condition. The number of HepG2 cells crossing Matrigel under acid condition was more than that under alkaline condition Under acidic conditions, the mRNA expression of EMT-related genes Vimentin, Slug, Snail and Zeb1 and the expression intensity of EMT-related proteins Vimentin and MMP9 in HepG2 cells were significantly higher than those in alkaline conditions. Conclusion Acidic environment can induce EMT in HepG2 cells.