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目的:探讨肝癌细胞中NFAT2基因启动子CpG岛甲基化状态及与其mRNA表达的关系。方法:用重硫酸盐测序法检测肝癌组织与癌旁组织及不同肝细胞系与正常肝细胞中NFAT2启动子甲基化状态,用qRT-PCR检测肝癌组织与癌旁组织NFAT2 mRNA表达,并分析肝癌组织NFAT2启动子甲基化与mRNA水平的相关性。结果:肝癌组织中NFAT2基因CpG岛甲基化率明显高于癌旁组织(33.0%vs.21.6%,P=0.003);人肝癌细胞系HuH7、HepG2、Hep3B中NFAT2基因CpG岛甲基化率分别为34.8%、40.4%、37.0%,均明显高于人正常肝细胞系L02中NFAT2基因CpG岛甲基化率(16.2%)(均P<0.05)。肝癌组织NFAT2 mRNA相对表达量明显低于与癌旁组织(0.000 602 4 vs.0.001 469,P<0.05),肝癌组织中NFAT2 mRNA水平与其启动子甲基化程度呈明显负相关(r=-0.661,P=0.027)。结论:肝癌细胞中NFAT2基因表达下调可能与其启动子区CpG岛高甲基化状态有关
Objective: To investigate the relationship between methylation status of NFAT2 promoter CpG island and its mRNA expression in hepatocellular carcinoma cells. Methods: The methylation status of NFAT2 promoter in hepatocellular carcinoma tissues, adjacent hepatocellular carcinoma tissues and normal hepatocytes was detected by bisulfite sequencing. The expression of NFAT2 mRNA was detected by qRT-PCR in hepatocellular carcinoma tissues and paracancerous tissues Relationship between NFAT2 promoter methylation and mRNA level in hepatocellular carcinoma. Results: The methylation rate of NFAT2 gene CpG island in HCC tissues was significantly higher than that in paracancer tissues (33.0% vs.21.6%, P = 0.003). The methylation rates of NFAT2 gene CpG island in human hepatocellular carcinoma cell lines HuH7, HepG2 and Hep3B (34.8%, 40.4%, 37.0%, respectively) were significantly higher than that of CpG island of NFAT2 gene in human normal liver cell line L02 (16.2%) (all P <0.05). The relative expression level of NFAT2 mRNA in hepatocellular carcinoma was significantly lower than that in para-cancerous tissues (0.000 602 4 vs.00.001 469, P <0.05). There was a negative correlation between the NFAT2 mRNA and the degree of methylation in hepatocellular carcinoma (r = -0.661 , P = 0.027). Conclusion: The down-regulation of NFAT2 gene expression in hepatocellular carcinoma may be related to the hypermethylation status of CpG island in its promoter region