CAJ | 학술논문

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150 nmol/L,低叶酸浓度为25 nmol/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.
목적 통과갑기화심편대협산결핍도치인정상결장점막상피세포(NCM460)적DNA갑기화상태변화진행사사,탐토상관위점여산발성결직장암발생적관계.방법 차조NimbleGen공사적갑기화심편,검측불동협산농도(고협산농도위150 nmol/L,저협산농도위25 nmol/L)배양적NCM460세포중기인계동자구CpG도적갑기화상태,병통과결합중아류산염적측서방법재세포화15례(남∶8,녀∶7)결직장암환자적조직양본중대심편적결과진행험증.결과 종고협산농도배양적NCM460세포중사선도료2680개발생료갑기화적CpG도,종저협산조사선도료3073개.재21호염색체상,17개기인계동자구CpG도발생료초갑기화혹거갑기화적개변,기중8개기인갑기화상태적개변가능여결직장암적발생상관.결론 협산결핍도치인NCM460적DNA갑기화상태개변가능여결직장암적발생상관.
Objective To investigate the changes in methylation patterns induced by folate deficiency 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 highresolution 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 cells 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 folate).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 NCM460 cells.Some related genes play roles in tumorigenesis of CRC.