MSRE-PCR法检测卵巢癌组织中OPCML基因甲基化  

Detection of OPCML methylation in tissues of ovarian cancer by MSRE-PCR

作　　者：周峰[1] 曹兴建[1] 马敏[1] 刘曼华[2] 陶国华[1] 

机构地区：[1]南通大学第二附属医院检验科,江苏南通226001 [2]南通大学第二附属医院妇产科,江苏南通226001

出　　处：《临床检验杂志》2011年第6期427-429,共3页Chinese Journal of Clinical Laboratory Science

基　　金：南通市社会发展科技项目(S2009017)

摘　　要：目的建立甲基化敏感性限制性内切酶-PCR(MSRE-PCR)法检测OPCML基因,探讨其在卵巢癌中的意义。方法用亚硫酸氢钠修饰测序法检测5例卵巢癌组织及正常卵巢组织中OPCML基因甲基化,分别用MSRE-PCR和巢式甲基化特异性PCR(MSP)检测组织DNA中OPCML基因甲基化发生率。结果 DNA序列中存在6个Bsh1236I酶切位点,酶切24 h能完全消化非甲基化DNA,MSRE-PCR和巢式MSP检测卵巢癌组织中OPCML基因甲基化发生率分别为86.7%和84.4%,正常卵巢组织两法均阴性。结论 MSRE-PCR检测卵巢癌组织OPCML基因甲基化发生率与巢式MSP相比没有显著性差异,是一种操作简便、敏感的甲基化检测方法。Objective To establish the method of methylation-sensitive restriction enzyme-PCR（MSRE-PCR） and investigate the significance of detection of opioid-binding cell adhesion molecule（OPCML） methylation in ovarian cancer.Methods MSRE-PCR for OPCML was established on the base of the known OPCML methylation status.The methylation rates of OPCML in the cancer tissues and normal tissues of 5 cases with ovarian cancer were detected by bisulfite sequencing method and the established MSRE-PCR.Results Six Bsh1236I digestion sites in the DNA sequence（243 bp） of OPCML were recognized,the DNA of non-methylation could be completely digested after the reaction with Bsh1236I for 24 hours.The methylation rates of OPCML in ovarian cancer tissues detected by MSRE-PCR and nested methylation-specific PCR（MSP） were 86.7% and 84.4% respectively,while no methylation of OPCML in normal ovarian tissue was detectable by the both methods.Conclusion The methylation rate of OPCML by MSRE-PCR was similar to that of MSP without statistically significant difference.MSRE-PCR should be an easy and sensitive method for monitoring methylation status of OPCML.

关 键 词：甲基化敏感性限制性内切酶 聚合酶链反应 结合鸦片样物质的细胞黏附分子基因 卵巢癌 

分 类 号：R737.3[医药卫生—肿瘤]