IL-6介导M2型巨噬细胞促进肝癌HepG2细胞上皮间充质样改变

IL-6 mediated M2 macrophages promote epithelial mesenchymal changes in HepG2 hepatoma cells

目的探索M2型巨噬细胞及其分泌因子白介素6(IL-6)对HepG2肝癌细胞上皮间充质样改变的影响。方法取人肝癌细胞株HepG2及由THP-1培养诱导成熟的M2型巨噬细胞、IL-6抑制剂Corylifol A进行实验。实验分两部分进行,第一部分设实验组(HepG2细胞株与M2型巨噬细胞共培组)及空白对照组(HepG2无特殊处理组);第二部分用HepG2细胞株与M2型巨噬细胞共培,设实验组及对照组,分别加入Corylifol A、无义溶媒。Western blot检测两组人肝癌细胞上皮间充质样表型转变指标E-钙黏蛋白(E-Cadherin)、N-钙黏蛋白(N-Cadherin)的蛋白表达水平。ELISA测定两组上清液IL-6表达水平。结果第一部分实验,与对照组相比,实验组E-Cadherin的mRNA表达及蛋白表达下降,M2型巨噬细胞可以抑制HepG2的E-Cadherin表达;与对照组相比,实验组N-Cadherin的mRNA及蛋白表达升高,M2型巨噬细胞可以促进HepG2的N-Cadherin表达。第二部分,与对照组相比,实验组E-Cadherin的蛋白表达升高,N-Cadherin的蛋白表达下降,M2型巨噬细胞分泌的IL-6抑制HepG2的E-Cadherin表达及促进N-Cadherin表达作用可被IL-6抑制剂抑制。结论IL-6介导M2型巨噬细胞促进肝癌HepG2细胞上皮间充质样改变。

Objective To investigate the effects of M2 macrophages and their secretory factor interleukin-6(IL-6)on epithelial mesenchymal changes in HepG2 hepatoma cells.Methods This study selected human hepatoma cell line HepG2,mature M2 macrophages induced by THP-1 culture and IL-6 inhibitor Corylifol A for the experiment.The experiment was divided into two parts.In the first part,the experiment group(HepG2 cell line and M2 macrophage co-culture group)and the control group(HepG2 without special treatment group)were set up.In the second part,HepG2 cell line was co-cultured with M2 macrophages,and the experiment group and the control group were added with Corylifol A and nonsense solvent respectively.The protein expression levels of E-Cadherin and N-Cadherin,indicators of epithelial mesenchymal phenotypic transformation,were detected by Western blot,and the expression level of IL-6 in supernatant of the two groups was determined by ELISA.Results In the first part of the experiment,compared with those of the control group,the mRNA and protein expression of E-Cadherin in the experiment group decreased.M2 macrophages could inhibit the expression of E-Cadherin in HepG2;compared with those of the control group,the mRNA and protein expression of N-Cadherin in the experiment group increased.M2 macrophages could promote the expression of N-Cadherin in HepG2.In the second part,compared with those of the control group,the protein expression of E-Cadherin increased and the protein expression of N-Cadherin decreased in the experiment group.IL-6 secreted by M2 macrophages could inhibit E-Cadherin expression and promote N-Cadherin expression in HepG2,which can be inhibited by IL-6 inhibitors.Conclusion IL-6 mediates M2 macrophages to promote epithelial mesenchymal changes of HepG2 hepatoma cells.