子宫颈癌SOX9基因甲基化早期诊断价值探讨

目的检测宫颈癌和宫颈上皮内瘤变（cervical intraepithelial neoplasia,CIN）脱落细胞中SOX9基因甲基化水平,为子宫颈癌的早期诊断提供新的分子标志。方法收集2010-03-01-2011-06-30就诊于首都医科大学附属北京妇产医院肿瘤科患者的宫颈脱落细胞156例,采用双探针实时定量聚合酶链式反应（polymerase chain reaction,PCR）技术,定量检测正常宫颈组（48例）、CINⅠ组（30例）、CINⅡ-Ⅲ组（30例）和宫颈癌组（48例）的宫颈脱落细胞中SOX9基因甲基化水平,分析SOX9基因甲基化与各级别宫颈病变的关系。结果正常宫颈组SOX9基因甲基化水平（甲基化评分）17.93±4.33,CINⅠ组为10.35±4.06,CINⅡ-Ⅲ组为33.17±15.19,宫颈癌组为48.04±24.03。SOX9基因甲基化水平随宫颈病变级别的增加而升高,且在CINⅡ-Ⅲ组和宫颈癌组中显著升高。CINⅠ组和正常宫颈组脱落细胞甲基化水平差异无统计学意义,P=0.062;SOX9基因甲基化水平在其余任意两组间差异均有统计学意义,P〈0.001。SOX9基因甲基化评分可有效区分宫颈癌/CINⅡ-Ⅲ和CINⅠ/正常宫颈,受试者工作特征曲线下面积（area under the receiver operating characteristic curve,AUC）为0.961（95%CI：0.933-0.990）,P〈0.001。在最佳临界值,SOX9基因甲基化评分检测宫颈癌/CINⅡ-Ⅲ的敏感度为92.3%,特异度为89.7%。结论甲基化SOX9基因能够有效诊断宫颈癌/CINⅡ-Ⅲ,可以作为子宫颈癌早期诊断的分子标志。

Investigation of SOX9 gene methylation in early diagnosis of cervical cancer

OBJECTIVE To detect the methylation level of SOX9 gene in cervical cancer and intraepithelial neoplasia scrapings and to provide new biomarker for early diagnosis of cervical cancer.METHODS Totally 156 cervical scrapings from patients of Beijing Obstetrics and Gynecology Hospital were collected.A TaqMan-based real-time PCR assay was carried out to detect the methylation level of the SOX9 gene in cervical scrapings,including 48 normal cervical scrapings,30 scrapings with cervical intraepithelial neoplasia 1（CINⅠ）,30 scrapings with CINⅡ-Ⅲ and 48 scrapings with cervical cancer.RESULTS The methylation level（methylation score）of the SOX9 gene in normal group was 10.35±4.06,in CINⅠ group 10.35±4.06,in CINⅡ-Ⅲ group 33.17±15.19,and in cervical cancer group 48.04±24.03.The methylation level increased significantly with the severity of cervical squamous lesions,a marked increase was observed in CINⅡ-Ⅲ and cervical cancer group,especially.No statistically significant difference was observed between normal and CINⅠ,P=0.062;however,the difference in methylation level was statistically significant between any other groups,P〈0.001.The methylation score of the SOX9 gene could be used to segregate cervical cancer/CINⅡ-Ⅲfrom CINⅠ/normal,with an AUC value of 0.961（95%CI,0.933-0.990）,P〈0.001.At the optimal cutoff value,a sensitivity of 92.3% and a specificity of 89.7% were obtained.CONCLUSIONS SOX9 methylation could diagnose SCC/CINⅡ-Ⅲ,and may provide a valuable molecular biomarker for early diagnosis of cervical cancer.