肝星状细胞对肝癌细胞株MHCC-LM3上皮间质转化的影响

Effect of hepatic stellate cell on epithelial-mesenchymal transition in hepatocellular carcinoma MHCC-LM3 cell line

目的探讨肝星状细胞LX-2对肝癌细胞株MHCC-LM3上皮间质转化(EMT)的影响。方法制备肝星状细胞LX-2上清,用LX-2上清和MHCC-LM3共培养(实验组),以单独培养的MHCC-LM3为对照(对照组),比较2组的肝癌细胞形态学、侵袭能力、EMT相关的上皮标记物E-钙黏蛋白(E-cadherin)及间质标记物波形蛋白Vimentin蛋白表达的变化。结果与对照组相比,实验组的肝癌细胞MHCC-LM3形态拉长,细胞间连接变得疏松。Transwell实验显示,实验组在共培养12、24、48 h的侵袭细胞数均比对照组增多(P均<0.01);随着培养时间的延长,实验组的侵袭细胞也随之增多(P均<0.01)。实验组MHCC-LM3细胞在共培养24、48 h时的E-cadherin蛋白表达水平均低于对照组细胞(P均<0.01),在共培养12、24、48 h时的Vimentin蛋白表达水平均高于对照组(P均<0.01)。结论肝星状细胞能够促进肝癌细胞株MHCC-LM3的EMT过程,增强其侵袭能力。

Objective To investigate the effect of hepatic stellate cells on the epithelial-mesenchymal transition （EMT）of hepatocellular carcinoma MHCC-LM3 cell line. Methods MHCC-LM3 cells were co-incubated with the culture supernatant of LX-2 in the study group,and the untreated MHCC-LM3 cells were as the control. The morphological changes,invasion ability,and the changes in the expression of E-cadherin and Vimentin proteins were statistically compared between two groups. Results Compared with the control group, the MHCC-LM3 cells co-incubated with the supernatant of LX-2 were elongated and the cell-cell junction became looser. Transwell assay revealed that the quantity of invasive MHCC-LM3 cells co-incubated for 12-,24-and 48-h in the study group was significantly higher compared with that in the control group （all P〈0.01 ）.Meantime,the quantity of invasive MHCC-LM3 cells in the study group was significantly enhanced over the time of incubation （all P〈0.01）. In the study group,the expression levels of E-cadherin protein after 24-and 48-h incubation were considerably lower than those in the control group （both P〈0.01）. The expression levels of Vimentin protein after 12-,24- and 48- incubation in the study group were significantly up-regulated compared with those in the control group （all P〈0.01）. Conclusion Hepatic stellate cells can promote the EMT and enhance the invasion of hepatocellular carcinoma MHCC-LM3 cell line.