Li Jian-Liang, Fei Qi, Yu Jian, Zhang Hong-Yu, Wang Peng, Zhu Jing-De
State Key Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai Jiaotong University, Shanghai,200032, PR China.
Ai Zheng. 2004 Sep;23(9):985-91.
BACKGROUND & OBJECTIVE: DNA methylation has been regarded as an important epigenetic signature reflecting the transcription state of DNA in cells. This study was to assess the correlation between methylation state of promoter CpG islands of metastasis-associated genes and their expression in 6 liver cell lines, including 5 cancerous.
Methylation specific polymerase chain reaction method (MSP) and DNA sequencing verification were used to analyze the methylation 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, HepG2, and HCCLM3), and 1 immortalized liver cell line (L-02). Expression of 6 genes in this list was assessed by the semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method.
The methylation state of genes was either unmethylated or heterozygously methylated in these 7 liver cell lines. Except for no expression of OXCT gene was detected by RT-PCR in both HepG2 and HCCLM3 cells where it was heterozygously methylated, there was expression of genes in all the remaining cases.
Although expression state of genes in this study supported the general notion that hypermethylation state of promoter 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.
DNA甲基化被视为反映细胞中DNA转录状态的重要表观遗传特征。本研究旨在评估转移相关基因启动子CpG岛的甲基化状态与其在6种肝细胞系(包括5种癌细胞系)中的表达之间的相关性。
采用甲基化特异性聚合酶链反应法(MSP)和DNA测序验证,分析7个基因(ASPH、ENO3、ITGA9、LRP6、MTHFD2、OXCT和SRP72)启动子CpG岛在5种肝癌细胞系(BEL-7402、SMMC-7721、Hep3B、HepG2和HCCLM3)及1种永生化肝细胞系(L-02)中的甲基化状态。通过半定量逆转录聚合酶链反应(RT-PCR)法评估该列表中6个基因的表达。
这7种肝细胞系中基因的甲基化状态为未甲基化或杂合甲基化。除了在杂合甲基化的HepG2和HCCLM3细胞中通过RT-PCR未检测到OXCT基因表达外,其余所有情况下基因均有表达。
尽管本研究中基因的表达状态支持基因启动子CpG岛的高甲基化状态代表基因转录沉默状态这一普遍观点,但也存在例外情况。因此,其他机制可能对本研究中这7个基因的观察到的表达状态有贡献。