肝癌细胞系中肝癌转移相关候选基因启动子区域CpG岛甲基化和表达水平的相关性研究(英文)  

Correlation between Methylation Profile of Promoter CpG Islands of Seven Metas tasis-associated Genes and Their Expression States in Six Cell Lines of Liver Origin

作　　者：李建良[1] 费琦[1] 余坚[1] 张红宇[1] 王鹏[1] 朱景德[1] 

机构地区：[1]上海交通大学上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《癌症》2004年第9期985-991,共7页Chinese Journal of Cancer

基　　金：中国高技术研究发展计划(863项目)(No.2002AA2Z3352);上海市科委基础研究项目(No.02DJ14056)~~

摘　　要：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 &OBJECTIVE: DNA methylation has been regarded as an im po rtant epigenetic signature reflecting the transcription state of DNA in cells. T his study was to assess the correlation between methylation state of promoter Cp G islands of metastasis-associated genes and their expression in 6 liver cell l ines, including 5 cancerous. METHODS: Methylation specific polymerase chain reac tion method (MSP) and DNA sequencing verification were used to analyze the methy lation state of promoter CpG islands of 7 genes (ASPH, ENO3, ITGA9, LRP6, MTHFD2 , OXCT, and SRP72) in 5 liver cancer cell lines (BEL-7402, SMMC-7721, Hep3B, H epG2, and HCCLM3), and 1 immortalized liver cell line (L-02). Expression of 6 g enes in this list was assessed by the semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method. RESULTS: The methylation state of ge nes was either unmethylated or heterozygously methylated in these 7 liver cell l ines. Except for no expression of OXCT gene was detected by RT-PCR in both HepG 2 and HCCLM3 cells where it was heterozygously methylated, there was expression of genes in all the remaining cases. CONCLUSION: Although expression state of ge nes in this study supported the general notion that hypermethylation state of pr omoter CpG islands of genes represents the silenced state of gene transcription, there were exceptions. Therefore, other mechanisms are likely to contribute to the observed expression state of these 7 genes in this study.

关 键 词：肝细胞肝癌 转移 启动子区域CpG岛 甲基化 甲基化特异性PCR 

分 类 号：R735.7[医药卫生—肿瘤] Q754[医药卫生—临床医学]