结直肠肿瘤DNA甲基化标志的筛选及诊断价值  

Screening and diagnostic value of the molecular markers of DNA methylation in colorectal neoplasma

作　　者：马炯[1] 杨青兰[4] 邓超[3] 金黑鹰[2] 龚伟[1] 王水明[2] 陈豪[3] 范怡梅[4] 

机构地区：[1]江苏省江阴市中医院外科,214400 [2]南京中医药大学第三附属医院全国肛肠医疗中心全国肛肠医疗中心江苏省中西医结合结直肠癌诊疗中心 [3]南京中医药大学第一临床医学院 [4]南京大学医学遗传教研室

出　　处：《中华胃肠外科杂志》2015年第11期1149-1153,共5页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金资助课题（81273944、30973837）;江苏省“六大人才高峰”高层次人才项目资助（2012-WS-087）;南京市“十二五”医学发展重大项目;南京市科技局社会发展项目（201402041）;南京市卫生局科技项目（YKK14140）

摘　　要：目的筛选有潜在诊断价值的甲基化基因，探讨其在国人结直肠肿瘤中的甲基化特征，以期寻找有较高诊断价值的DNA甲基化筛查标志。方法收集2013年6—8月期间于南京中医药大学第三附属医院接受手术治疗的10例结直肠癌患者的肿瘤及其距离肿瘤10cm以上组织标本及接受内镜下切除的10例结直肠腺瘤患者的组织标本。分别采用BSP法和qMSP法检测上述标本中SNCA、MAL、INA、SPG20、FBN1、CNRIP1、TFP12和0sMR8个基因的甲基化特征，筛选出有潜在筛查价值的基因，并绘制接受者工作特征（ROC）曲线。结果BSP法检测的8个基因中，SNCA、SPG20和FBN1基因在癌和腺瘤组织中100％检测到甲基化，而在癌旁组织中均未检测到甲基化：其余5个基因在癌、癌旁和腺瘤中均有不同程度表达。qMSP法检测的10对癌与癌旁组织中的SNCA甲基化阳性例数分别为5例和1例，SPG20分别为8例和1例，FBN1分别7例和0例，差异均有统计学意义（分别P=0．070、P=0．003和P=0．007）。SNCA、SPG20和FBN1基因甲基化诊断结直肠癌的ROC曲线下面积（AUC）分别为0．890、0．730和0．880。结论SNCA、SPG20和FBN1对于结直肠肿瘤为具有潜在筛查价值的基因。Objective To screen the molecular markers of DNA methylation with potential diagnostic value, and to explore their methylation features in Chinese colorectal neoplasma in order to find out ones with higher diagnostic value. Methods Tissue samples of colorectal cancer and normal adjacent mucosa （〉10 cm distance to tumors） from 10 colorectal cancer patients undergoing operation, and tissue samples of colorectal adenoma from 10 patients undergoing endoscopic resection in our center from June to August 2013 were collected respectively. Methylation status of 8 genes, such as SNCA, MAL, INA, SPG20, FBN1, CNRIP1, TFPI2, OSMR, was detected by BSP and qMSP to screen genes with potential diagnostic valua. ROC curve was drawn to analyze its diagnostic value. Results BSP measurement showed that the rate of DNA methylation of SNCA, SPG20 and FBN1 was 100% in colorectal cancer and adenoma, while no methylation was found in normal adjacent mucosa. The other 5 genes expressed in different extent in cancer, adenoma and normal adjacent mucosa. Among 10 cancer tissues and normal adjacent mucosa detected by qMSP method, positive SNCA methylation was found in 5 cases and 1 case respectively; positive SPG20 in 8 cases and 1 case respectively; positive FBN1 in 7 cases and 0 cases respectively, whose differences were significant （P=0.070, P=0.003 and P=0.007）. The area under curve （AUC） of SNCA, SPG20, and FBN1 methylation for diagnosing colorectal cancer was 0.890, 0.730 and 0.880 respectively. Conclusion SNCA, SPG20 and FBN1 are potential genes with screening value for colorectal neoplasma.

关 键 词：结直肠肿瘤 基因 甲基化 肿瘤筛查 

分 类 号：R735.34[医药卫生—肿瘤]