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目的:检测Sall3基因在肝癌细胞系和肝癌组织中的甲基化状况及其表达水平。方法:对7株肝癌细胞系、142对肝癌组织和6例正常肝组织进行Sall3基因启动子区域甲基化检测。甲基化研究使用甲基化特异性聚合酶链反应(methylation specific polymerase chain reaction,MSP),基因表达水平研究采用实时定量反转录PCR(real-time RT-PCR)。结果:MSP结果显示,7株肝癌细胞株中有6株Sall3基因的启动子区域为纯合甲基化,1株为纯合去甲基化;在6例正常肝组织中Sall3均为纯合去甲基化;在142对肝癌组织标本中,癌旁组织Sall3基因启动子区域甲基化率为18.31%,在癌组织中为64.79%。real-time RT-PCR结果显示,肝癌肿瘤细胞系Sall3mRNA的表达水平明显低于去甲基化的正常肝组织。结论:Sall3基因启动子甲基化在正常肝组织、肝癌组织(和肝癌细胞系)中有明显差异。
Objective: To detect the methylation and expression of Sall3 gene in hepatocellular carcinoma and hepatocellular carcinoma. Methods: The methylation status of Sall3 gene promoter in 7 hepatocellular carcinoma cell lines, 142 hepatocellular carcinoma tissues and 6 normal liver tissues was detected. Methylation-specific methylation-specific polymerase chain reaction (MSP) was used for methylation analysis and real-time RT-PCR for gene expression analysis. Results: The results of MSP showed that the promoter region of 6 Sall3 genes in 7 hepatoma cell lines was homozygous methylation and 1 was homozygous demethylation. Sall3 was homozygous in 6 normal liver tissues In 142 pairs of HCC tissues, the methylation rate of Sall3 gene in promoter region was 18.31% in cancer tissues and 64.79% in cancer tissues. Real-time RT-PCR results showed that the expression level of Sall3 mRNA in hepatocellular carcinoma cell line was significantly lower than that in normal demethylated liver tissue. CONCLUSIONS: Sall3 promoter methylation is significantly different in normal liver tissue and liver cancer tissue (and in hepatoma cell line).