摘要
目的通过甲基化芯片对叶酸缺乏导致人正常结肠黏膜上皮细胞（NCM460）的DNA甲基化状态变化进行筛查，探讨相关位点与散发性结直肠癌发生的关系。方法借助NimbleGen公司的甲基化芯片，检测不同叶酸浓度（高叶酸浓度为150nmol／L，低叶酸浓度为25nmol／L）培养的NCM460细胞中基因启动子区CpG岛的甲基化状态，并通过结合重亚硫酸盐的测序方法在细胞和15例（男：8，女：7）结直肠癌患者的组织样本中对芯片的结果进行验证。结果从高叶酸浓度培养的NCM460细胞中筛选到了2680个发生了甲基化的CpG岛，从低叶酸组筛选到了3073个。在21号染色体上，17个基因启动子区CpG岛发生了超甲基化或去甲基化的改变，其中8个基因甲基化状态的改变可能与结直肠癌的发生相关。结论叶酸缺乏导致人NCM460的DNA甲基化状态改变可能与结直肠癌的发生相关。
Objective To investigate the changes in methylation patterns induced by folate defi- ciency in NCM460 cells and the correlation between the related loci and sporadic colorectal cancer （CRC）. Methods NimbleGen MeDIP chip （Methylated DNA Immunoprecipitation chip） assay was used in high- resolution mapping of DNA methylation patterns in NCM460 cells cultured with 150 and 25 nmol/L folate respectively. The results were verified by bisulfate sequencing PCR （BSP） in NCM460 ceils and specimens from 15 patients （8 males and 7 females） with sporadic CRC. Results The number of methylated CpG islands was 2680 （cells cultured with 150 nmol/L folate） and 3073 （cells cultured with 25 nmol/L fo- late）. There were 17 genes with hyper- or hypomethylation in the CpG island of promoter on chromosome 21 and 8 of them seemed to be associated with tumorigenesis of CRC. Conclusion Folate deficiency could induce aberrant DNA methylation patterns in NCMd60 cells. Some related genes play roles in tumorigenesis of CRC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第2期191-194,共4页
Chinese Journal of Experimental Surgery
