摘要
目的:探讨粪便miR34b/c甲基化状态的检测在结直肠癌早期诊断中的意义.方法:从126例结直肠癌患者癌组织、癌旁组织、粪便和64例正常对照者的粪便中分别提取DNA,采用多重置换扩增(multiple displacement amplification,MDA)技术对经过亚硫酸氢盐修饰样本进行全基因组扩增,结合甲基化特异性PCR(methylation-specific PCR,MSP)检测组织和粪便中miR34b/c基因甲基化状态.结果:结直肠癌癌组织miR34b/c基因的甲基化阳性率为95.2%(120/126),对应的癌旁正常组织为11.9%(15/126),两者比较有显著差异(P<0.01);miR34b/c甲基化状态与各临床病理参数无显著相关(P>0.05).结直肠癌粪便miR34b/c甲基化阳性率为90.2%(111/123),显著高于正常对照7.8%(5/64),差异有统计学意义(P<0.01).粪便DNAmiR34b/c用于结直肠癌早期诊断的敏感性为91.2%,特异性为92.2%.结论:miR34b/c甲基化是结直肠癌的重要分子特征,检测粪便miR34b/c甲基化有望成为结直肠癌早期诊断的一个全新的肿瘤标志物.MDA结合MSP为miRNA的甲基化分析提供了一种较理想的研究手段.
AIM： To investigate the value of detection of stool miR34b/c methylation in the diagnosis of colorectal cancer. METHODS： Multiple displacement amplification （MDA） was used to amplify bisulfite modi- fied genomic DNA, and methylation-specific PCR （MSP） was used to analyze methylation of miR34b/c in colorectal cancer tissue and stool DNA from 126 patients with colorectal cancer and stool DNA in 64 patients with benign diseases. RESULTS： In 126 cancer specimens and matched tumor-adjacent tissue specimens, 95.2% （120/126） and 11.9% （15/126） showed methylation of miR34b/c, and there is a signifi- cant difference in the rate of methylation be- tween them （P 〈 0.01）. There was no significant correlation between methylation of miR34b/c and clinicopathologic parameters （all P 〉 0.05）. The rate of methylation of miR34b/c in stool DNA was significantly higher in cancer patients than in patients with benign disease （90.2% vs 7.8%, P 〈 0.01）. The sensitivity and specificity of detection of miR34b/c methylation in diagnosis of colorectal cancer were 90.2% and 92.2%, re- spectively. CONCLUSION： The hypermethylation of miR34b/c is frequent in colorectal cancer and may be used as a novel diagnostic biomarker for colorectal cancer. MDA and MSP techniques provide ideal tools for analysis of methylation in trace DNA samples.
出处
《世界华人消化杂志》
CAS
北大核心
2013年第8期724-728,共5页
World Chinese Journal of Digestology
基金
解放军总医院苗圃基金资助项目
No.09KMM30~~
