宫颈癌SOX9基因甲基化水平的研究  

Identification of DNA methylation of SOX9 in cervical cancer using methylated-CpG island recovery array

作　　者：巫剑红[1] 梁学爱[1] 代荫梅[1] 

机构地区：[1]首都医科大学附属北京妇产医院微创中心,北京100026

出　　处：《中华肿瘤防治杂志》2013年第19期1513-1516,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：北京市教委科技计划面上项目(KM201010025022);北京市科技计划项目(D09050703570902);北京市科委重大项目(9050703560905)

摘　　要：目的:通过全基因组的甲基化芯片,筛选出宫颈癌新的甲基化基因,并在宫颈脱落细胞中验证其检测宫颈癌的应用价值。方法:采用CpG岛富集化分析甲基化芯片(methylated-CpG island recovery assay,MIRA)技术在宫颈癌和正常宫颈组织中筛选异常甲基化基因,选择甲基化水平较高的性别决定基因9(sex determining region Y-box 9,SOX9),应用亚硫酸氢钠-限制性酶切法在48例宫颈癌和48例正常宫颈脱落细胞中检测SOX9基因甲基化状态。结果:1)通过MIRA技术可确定宫颈癌组织中504个CpG岛,对应378个基因为异常甲基化基因,30个为显著甲基化基因,其中LMX1A和ZNF135已在宫颈癌中有所报道,TLX3、TLX2、PAX6、EN2、EFNA5、TBX5、SOX9、HOXB13、BCL11B、LHX2、DLX4、TLX1、PKLR和OLIG2已在其他肿瘤组织中发生甲基化,其余14个基因鲜见甲基化的相关报道。这30个基因中,SOX9为甲基化水平较高的基因。2)48例宫颈癌脱落细胞中32例发生SOX9甲基化,48例正常宫颈脱落细胞中未发现发生甲基化。宫颈癌脱落细胞中SOX9甲基化水平显著高于正常宫颈脱落细胞,差异有统计学意义,χ2=48.000,P=0.000。SOX9基因甲基化检测宫颈癌的敏感度为67%(32/48),特异度为100%(48/48)。结论:SOX9基因在宫颈癌中频繁发生甲基化,甲基化SOX9可能成为宫颈癌检测的分子标志。OBJECTIVE：To identify novel candidate methylation markers for cervical cancer detection,and then to validate the significantly hypermethylated genes in cervical scrapings.METHODS：MIRA-based array was carried out in dye swap way in cervical tissues to screen significantly hypermethylated genes in cervical cancer.COBRA assay in an independent set of 48cervical cancer and 48normal cervical scrapings was conducted to detect the methylation level of SOX9.RESULTS：Totally 504CpG islands,corresponding to 378genes,were identified to be differentially methylated between cervical cancer and normal cervical tissues,30genes were found to be significantly hypermethylated.Among them,LMX1Aand ZNF135had been previously reported as hypermethylated in cervical cancer.TLX3,TLX2,PAX6,EN2,EFNA5,TBX5,SOX9,HOXB13,BCL11B,LHX2,DLX4,TLX1,PKLR and OLIG2had been reported as hypermethylated in other cancers.Among the 30genes,SOX9had the highest methylation level.Methylated SOX9was detected in 0of 48controls and in 32of 48cervical cancers.SOX9methylation level in cervical cancer scrapings was significantly higher than that in normal cervical scrapings（χ2=48.000,P=0.000）.The sensitivity and specificity of SOX9gene methylation for cervical cancer detection was 67%（32/48）and 100%（48/48）,respectively.CONCLUSION：SOX9methylation is frequently involved in cervical carcinogenesis,and may provide a valuable marker for improved detection of cervical cancer.

关 键 词：宫颈肿瘤 DNA甲基化 染色体性别决定基因9 微阵列 

分 类 号：R737.33[医药卫生—肿瘤]