TGF-β1诱导肝星状细胞活化调控肝癌的炎症和免疫反应

TGF-β1 induced activation of hepatic stellate cells to regulate inflammation and immunoreaction in hepatocellular carcinoma

目的通过TGF-β1诱导活化的肝星状细胞与肝癌细胞SMMC7721共培养,观察肝癌的炎症和免疫变化。方法选取厦门大学附属福州第二医院体外培养人肝星状细胞LX-2,用10μg/L TGF-β1处理LX-224 h,取活化LX-2上清液共培养SMMC7721细胞12 h后,用Disitetide处理共培养细胞,流式细胞术观察CD4+CD25+T细胞干预前后的表达变化,ELISA观察白细胞介素-8(IL-8)、白细胞介素-10(IL-10)干预前后的表达情况。结果10μg/L的TGF-β1刺激LX-2后,α-SMA基因和蛋白水平表达均明显增加。活化的LX-2上清液与SMMC7721共培养后,CD_(4)^(+)CD_(25)^(+)的表达比例较SMMC7721中明显增加(P=0.003),IL-8和IL-10的表达水平也明显上调(P<0.001),用Disitetide抑制TGF-β1后CD_(4)^(+)CD_(25)^(+)的表达比例较共培养组中明显减少(P=0.025),IL-8和IL-10的表达水平也明显下调(P<0.001)。结论TGF-β1可以诱导肝星状细胞活化从而促进肝癌细胞的炎症和免疫反应。

Objective To observe the inflammatory and immune changes of hepatocellular carcinoma(HCC)by co-culture of activated hepatic stellate cells induced by TGF-β1 with hepatocellular carcinoma cells of SMMC7721.Methods Human hepatic stellate cells(LX-2)were cultured in Fuzhou No.2 Hospital Affiliated to Xiamen University and treated with 10μg/L TGF-β1 for 24 h.SMMC7721 cells were co-cultured with activated LX-2 supernatant for 12 h,and then treated with Disitetide.The expression changes of CD4+CD25+T cells before and after the intervention were observed by flow cytometry,and the expression of IL-8 and IL-10 before and after the intervention were observed by ELISA.Results After 10μg/L TGF-β1 stimulation of LX-2,α-SMA gene and protein expression were significantly increased.After co-culture of activated LX-2 supernatant with SMMC7721,the expression ratio of CD4+CD25+was significantly increased(P=0.003)compared with SMMC7721,and the expression levels of IL-8 and IL-10 were also significantly up-regulated(P<0.001).After TGF-β1 inhibition with Disitetide,the expression ratio of CD4+CD25+was significantly decreased(P=0.025),and the expression levels of IL-8 and IL-10 were also significantly down-regulated(P<0.001),compared with the co-culture group.Conclusion TGF-β1 can induce the activation of hepatic stellate cells and promote inflammation and immunoreaction in hepatocellular carcinoma cells.