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背景与目的:DNA甲基化被认为是反映细胞内DNA转录状态的重要表观遗传学标记。本研究旨在对启动子区域CpG岛的甲基化与基因转录水平的相关性进行评估。方法:采用甲基化特异性PCR的方法确定7个与肝癌转移相关的候选基因的启动子区域在6个肝细胞系(包括5个肝癌源性的)中的甲基化状态,并通过半定量PCR的方法确定6个基因稳定态的mRNA水平。结果:仅有纯合和杂合去甲基化的基因状态得到发现。RT-PCR分析表明除了OXCT基因在其处于杂合去甲基化状态的HepG2和HCCLM3细胞中不表达,在其他的情况下,7个候选基因均有表达。讨论:本研究中的7个肝癌转移相关基因的甲基化状态仅在细胞系中部分的反映了基因的转录状态,提示其它转录调控机制的存在。
BACKGROUND & AIM: DNA methylation is considered as an important epigenetic marker that reflects the transcriptional status of DNA in cells. The purpose of this study was to evaluate the association between promoter methylation and gene transcription at CpG island. METHODS: Methylation-specific PCR was used to determine the methylation status of seven promoter regions of candidate genes associated with liver cancer metastasis in six hepatocellular lines, including five hepatocarcinomas, Quantitative PCR was used to determine the steady-state mRNA levels of six genes. Results: Only the homozygous and heterozygous demethylated genes were found. RT-PCR analysis showed that except for OXCT genes that were not expressed in HepG2 and HCCLM3 cells that were heterozygous and demethylated, all seven candidate genes were expressed in other cases. Discussion: The methylation status of seven HCC metastasis-related genes in this study partially reflects the transcriptional status of genes in cell lines, suggesting the existence of other transcriptional regulatory mechanisms.