γ-干扰素对肝癌细胞增殖和抗原表达影响的相关研究

The related study on the effects of interferon-γ on cellular proliferation and antigens expression of hepatocellular carcinoma

目的探讨IFN-γ对肝癌细胞肿瘤抗原甲胎蛋白和MAGE-1表达的影响与机制。方法用重组人干扰素处理人肝癌细胞系SMMC-7721,用四氮唑比色法检测人肝癌细胞增殖活性,用间接免疫荧光检测MAGE-1表达,ELISA法检测甲胎蛋白表达,用BSP法测MAGE-1基因启动子甲基化状态,用免疫沉淀测JNK活性。结果在IFN-γ作用下,SMMC-7721细胞甲胎蛋白表达量下降,MAGE-1表达下降,MAGE-1基因启动子甲基化程度增加而JNK活性降低。结论 IFN-γ可增强MAGE-1基因启动子甲基化程度,下调MAGE-1表达,降低JNK活性,抑制细胞增殖。

Objective To investigate the effects and mechanisms of Interferon-γ（IFN-γ） on the expression of hepatocellular carcinoma-associated antigens AFP and MAGE-1. Methods Human hepatocellular carcinoma cell line SMMC-7721 cells were treated with recombinant IFN-γ.MTT colorimetry was used to reveal the proliferative activity.ELISA method was applied to detect the AFP expression.Indirect immunofluorescence was used to detect MAGE-1 expression.Bisulfite-sequencing PCR（BSP） was used to assay the methylation status of MAGE-1 gene promoter of SMMC-7721.Immunoprecipitation was used to assay the activity of the c-Jun N-terminal kinases. Results The results of indirect immunofluorescence showed that the expression of MAGE-1 and AFP in IFN-γ-treated SMMC-7721 were weaker than those in untreated cells.The methylation status of MAGE-1 gene promoter in IFN-γ-treated SMMC-7721 was higher than that in untreated cells.The activity of JNK decreased in IFN-γ-treated cells. Conclusion IFN-γ may enhance methylation status of MAGE-1 gene promoter,down-regulate the expression of MAGE-1 and decrease JNK activity.