CAJ | 학술논문

目的探讨胱硫醚-β-合酶(CBS)启动子区的超甲基化与基因表达及功能的相关性.方法通过结合重亚硫酸盐处理的直接测序、实时定量聚合酶链反应(PCR)、荧光偏振免疫法及免疫组织化学等技术对不同叶酸浓度( 25、150 nmol/L)下培养的人正常结肠黏膜上皮细胞株(NCM460)细胞和患者组织样本(包括肿瘤组织及相应的正常组织95例)中CBS基因启动子区CpG岛超甲基化与基因表达及功能进行研究.结果低叶酸浓度培养下NCM460细胞中CBS基因的表达下调,检测高甲基化组(H-M组:＞41.7％,83例)和低甲基化组(L-M组:≤41.7％,107例)组织样本中CBS基因表达水平,两组差异有统计学意义(P＜0.01).在NCM460细胞内,CBS基因的下调能够导致同型半胱氨酸积聚,检测H-mRNA组(＞3.19,45例)和L-mRNA组(≤3.19,40例)患者的血清中同型半胱氨酸的水平,L-mRNA组患者血清中同型半胱氨酸的水平显著高于H-mRNA组.结论 CBS启动子区的超甲基化极有可能导致CBS基因表达水平的下调,影响细胞内的甲基代谢,进而参与结直肠癌的发生、发展等一系列病程.
목적 탐토광류미-β-합매(CBS)계동자구적초갑기화여기인표체급공능적상관성.방법 통과결합중아류산염처리적직접측서、실시정량취합매련반응(PCR)、형광편진면역법급면역조직화학등기술대불동협산농도( 25、150 nmol/L)하배양적인정상결장점막상피세포주(NCM460)세포화환자조직양본(포괄종류조직급상응적정상조직95례)중CBS기인계동자구CpG도초갑기화여기인표체급공능진행연구.결과 저협산농도배양하NCM460세포중CBS기인적표체하조,검측고갑기화조(H-M조:＞41.7％,83례)화저갑기화조(L-M조:≤41.7％,107례)조직양본중CBS기인표체수평,량조차이유통계학의의(P＜0.01).재NCM460세포내,CBS기인적하조능구도치동형반광안산적취,검측H-mRNA조(＞3.19,45례)화L-mRNA조(≤3.19,40례)환자적혈청중동형반광안산적수평,L-mRNA조환자혈청중동형반광안산적수평현저고우H-mRNA조.결론 CBS계동자구적초갑기화겁유가능도치CBS기인표체수평적하조,영향세포내적갑기대사,진이삼여결직장암적발생、발전등일계렬병정.
Objective To explore the association between the hypermethylation of cystathioninebeta-synthase (CBS) promoter and gene expression and function.Methods Bisulfate sequencing polymerase chain reaction (PCR),real-time PCR,fluorescence polarization immunoassay and immunostaining were used to investigate the relationship between the hypermethylation of CpG island in CBS promoter and gene expression and function in NCM460 cultured with varying folate concentrations and patients' tissues.Results In the NCM460 cultured with low concentrations of folate,the expression of CBS was down-regulated,and there was significant difference in the gene expression of CBS between H-M group and L-M group (P ＜0.01 ).In NCM460 cells,the results represented that along with the down-regultaion of CBS,there was a significant accumulation of homocysteine,and the HCY level was higher in L-mRNA group than in H-mRNA group.Conclusion Hypermethylation of CBS promoter is likely to induce decrease of CBS gene expression,thereby it may influence intracellular methyl metabolism and involved in the occurrence and progression of colorectal cancer.