nMSP-DHPLC检测粪便SEPT9基因甲基化及其在结直肠癌诊断中的应用  

Detection of SEPT9 gene methylation by nMSP-DHPLC in stool DNA and its employment in early diagnosis of colorectal cancer

作　　者：赵慧霞[1] 李秋文[1] 董伟伟[1] 段昕妤[1] 朱建华[1] 王如良[1] 郝怡鑫[1] 叶明[1] 肖文华[1] 

机构地区：[1]解放军总医院第一附属医院肿瘤科,北京100048

出　　处：《军事医学》2012年第5期388-391,共4页Military Medical Sciences

基　　金：解放军总医院苗圃基金资助(09KMM30)

摘　　要：目的探讨粪便SEPT9(septin 9)基因甲基化检测在结直肠癌早期诊断中的应用价值。方法常规提取87例结直肠癌手术切除癌组织、癌旁正常结肠组织及其术前粪便DNA,应用巢式甲基化特异性PCR(nested methyl-ation specific PCR,nMSP)结合变性高效液相色谱(denatured high performance liquid chromatography,DHPLC)技术检测结直肠癌组织、癌旁组织及粪便SEPT9基因甲基化水平;比较结直肠癌组织和粪便中SEPT9甲基化检测在结直肠癌诊断中的敏感性和特异性。结果在87例结直肠癌中,癌组织SEPT9检出率为80.5%(70/87),癌旁组织检出率为8.0%(7/87),二者比较差异有统计学意义(P<0.01),特异性为91.9%;结直肠癌粪便SEPT9基因甲基化检测结果与组织检测结果基本一致。SEPT9基因甲基化与大于65岁老年患者和右半结肠癌显著相关。结论结直肠癌组织SEPT9基因甲基化异常发生率高,尤其是老年和右半结肠癌患者;粪便SEPT9基因甲基化异常可作为结直肠癌早期诊断的肿瘤标志物。Objective To investigate the value of analyzing septin（SEF19） gene methylation in stool DNA for diagnosis of colorectal cancer. Methods The colorectal carcinoma, para-careinoma tissues,and stool SEPT9 gene methylation were detected by nested methylation specific PCR（nMSP） arid denatured high performance liquid chromatography （DHPLC） in 87 colorectal cancer cases. The sensitivity and specificity of SEPT9 methylation marker for coloreetal cancer diagnosis were analyzed. Results Among 87 colorectal cancer cases, SEPT9 gene methylation was found in 80.5% （ 70/87 ） of eolorectal carcinoma tissues and 8.0% （7/87） of para-carcinoma tissues, respectively. The sensitivity and specificity of SEF19 were 80.5% and 91.9% ,respectively, and the stool DNA was almost as sensitive as cancer tissues in diagnosis of coloreetal cancer. The methylation of SEPT9 was significantly associated with elders above 65 years old and the right colon cancer. Conclusion Abnormal methylation of SElYP9 occurs frequently in colorectal cancer, especially in the elders and the right colon cancer, SEPT9 gene mehylation in stool DNA can be used as a candidate tumor marker for early diagnosis of colorectal cancer.

关 键 词：SEPT9/甲基化 粪便 DNA 结直肠肿瘤 色谱法 高压液相 诊断 

分 类 号：R735.3[医药卫生—肿瘤]