甲基化特异性PCR联合结合重亚硫酸盐限制性内切酶法在胃癌抑癌基因甲基化检测中的应用  

Application of combining MSP and COBRA for the detection of methylation of tumor suppressor gene in gastric cancer

作　　者：胡世莲[1,2] 程昭栋[1,2] 孙玉蓓[1,3] 徐维平[4] 沈干[1,2] 孔祥勇[1,2] 

机构地区：[1]安徽医科大学附属省立医院 [2]安徽省立医院老年医学科,合肥230001 [3]安徽省立医院肿瘤内科,合肥230001 [4]安徽省循证医学中心

出　　处：《中国临床保健杂志》2010年第2期113-116,F0003,共5页Chinese Journal of Clinical Healthcare

基　　金：国家自然科学基金资助项目(30672383)

摘　　要：目的探讨甲基化特异性PCR(MSP)方法检测胃癌组织中抑癌基因甲基化的准确性,联合应用MSP方法与结合重亚硫酸盐限制性内切酶法(COBRA)检测抑癌基因甲基化状态的意义。方法采用MSP检测胃癌组织抑癌基因Runx3启动子区甲基化的状态,随后再采用COBRA检测胃癌组织标本的甲基化状态。分析两种方法检测结果的关系。结果经MSP方法检测54例肿瘤组织中共有30例标本发生Runx3基因启动子区异常甲基化,甲基化阳性率为55.6%。经COBRA方法检测54例肿瘤组织中有27例出现酶切条带即存在甲基化,甲基化阳性率为50.0%。经COBRA方法检测,在MSP方法检测的30例阳性标本中有3例未出现酶切条带即为假阳性。在MSP方法检测Runx3基因甲基化阴性的24例标本中,经COBRA方法检测,Runx3基因甲基化阴性的标本亦为24例。两种检测方法检测结果的阳性率差异无统计学意义(P>0.05)。结论甲基化特异性PCR方法在检测胃癌抑癌基因甲基化状态的研究中具有较高的灵敏度,COBRA方法可以检测出MSP方法中的假阳性标本,联合应用两种方法检测胃癌组织中抑癌基因的甲基化状态会使实验结果更可靠、准确。Objective To investigate the accuration of detection for Methylation Specific PCR in tumor suppressor gene（TSG）of carcinoma specimens from gastric cancer.And study the significance of combine MSP and COBRA to detect the methylation of TSG in gastric cancer.Methods Firstly,MSP was used to detect the promoter methylation of Runx3 gene of specimens from gastric cancer,then we used COBRA to investigate the methylation of Runx3 gene from the same specimens.At last,we analyze the relationship between MSP and COBRA.Results The DNA methylation of the Runx3 gene was found in 30 of the 54 gastric cancer samples（55.6%）.By contrast,the positive rates of the same samples was 50.0%（27/54）,which detected by COBRA.3 methylated-positive and 24 methylated-negtive samples which detected by MSP were undigested by COBRA.No significant difference was found on the results between MSP and COBRA for detecting the aberrant methylation of promoter-associated CpG islands of CHFR gene in gastric cancer samples（P0.05）.Conclusion In our present study,MSP possesses high sensitivity for detecting the methylation on TSG.COBRA possess the effect of investigation on false positive of MSP.Combined MSP and COBRA will make the experimental result more reliable and precise for detecting the promoter methylation of TSG in gastric cancer.

关 键 词：胃肿瘤 DNA甲基化 聚合酶链反应 基因 肿瘤抑制 限制性内切酶图谱法 

分 类 号：R735.2[医药卫生—肿瘤]