论文部分内容阅读
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.
OBJECTIVE: To investigate the significance of fecal miR34b/c methylation status in the early diagnosis of colorectal cancer. Methods: From feces of 126 cases of colorectal cancer tissues, adjacent tissues, feces and 64 cases of normal controls DNA was extracted, and the bisulfite modified samples were subjected to whole genome amplification using multiple displacement amplification (MDA) technique, and methylation-specific PCR (MSP) was used to detect tissues and feces. Methylation status of miR34b/c gene. Results: The methylation positive rate of miR34b/c gene in colorectal cancer tissue was 95.2% (120/126), corresponding to adjacent normal tissues was 11.9% (15/126). There was a significant difference between the two groups (P<0.01); the miR34b/c methylation status was not significantly associated with any clinicopathological parameters (P>0.05). The positive rate of miR34b/c methylation in colorectal cancer feces was 90.2% (111). /123), significantly higher than the normal control 7.8% (5/64), the difference was statistically significant (P<0.01). The fecal DNA miR34b/c was used for the early diagnosis of colorectal cancer with a sensitivity of 91.2% and a specificity of 92.2. %. Conclusions: miR34b/c methylation is an important molecular feature of colorectal cancer. Detection of miR34b/c methylation in stools is expected to become A new tumor marker for early diagnosis of colorectal cancer was .MDA miRNA binding MSP methylation analysis provides an ideal means of research.