人乳头瘤状病毒阳性患者PAX1、TP63基因启动子甲基化状态检测及临床意义  

Detection and clinical significance of PAX1 and TP63 gene promoter methylation in HPV positive patients with different degrees of cervical lesions

作　　者：王慕喆 邢秀月 王丽娜 周叶 李晓丽 WANG Muzhe;XING Xiuyue;WANG Li'na;ZHOU Ye;LI Xiaoli(Department of Gynecology,Qionghai People's Hospital,Qionghai,Hainan 571400,China)

机构地区：[1]琼海市人民医院妇科,海南琼海571400

出　　处：《中国热带医学》2023年第12期1336-1340,共5页China Tropical Medicine

基　　金：琼海市科技计划项目(No.海科工信[2021]63)。

摘　　要：目的探究宫颈人乳头瘤状病毒(human papilloma virus,HPV)感染阳性患者宫颈病变程度与配对盒家族基因1(paired box-1,PAX1)、肿瘤蛋白63(tumor protein 63,TP63)基因启动子甲基化状态的关系,分析其检测的临床意义。方法收集2021年12月—2022年12月于琼海市人民医院就诊的128例疑似宫颈病变合并HPV感染阳性患者的宫颈组织细胞标本,根据病理检查结果分为低级别鳞状上皮内病变组(low grade squamous intraepithelial lesion,LSIL,n=43)、高级别鳞状上皮内病变(high grade squamous intraepithelial lesion,HSIL,n=51)和宫颈癌组(n=34)。采用第二代杂交捕获法检测HPV病毒定量,亚硫酸氢盐测序法检测PAX1、TP63基因启动子甲基化度,实时定量PCR法检测宫颈组织PAX1、TP63相关mRNA表达;评估PAX1、TP63甲基化程度对宫颈上皮内瘤样病变2+(cervical intraepithelial neoplasia 2+,CIN2+)的诊断效能。结果各组HR-HPV定量结果比较,差异有统计学意义(P>0.05)。128例患者中,共检出PAX1基因启动子甲基化49例(38.28%),TP63基因启动子甲基化55例(42.97%)。宫颈癌组PAX1、TP63基因启动子甲基化率分别为67.65%、73.53%,HSIL组分别为43.14%、49.02%,LSIL组分别为9.30%、11.63%,差异有统计学意义(P<0.05)。宫颈癌组PAX1、TP63 mRNA表达水平分别为(0.34±0.08)、(0.45±0.13),HSIL组分别为(0.72±0.11)、(0.63±0.09),LSIL组分别为(1.04±0.09)、(0.87±0.11),差异有统计学意义(P<0.05)。受试者工作特征(receiver operating characteristic,ROC)曲线分析结果显示,PAX1、TP63基因启动子甲基化及其联合检测诊断CIN2+病变的曲线下面积(area under curve,AUC)值分别为0.793、0.842和0.857,灵敏度分别为77.78%、83.33%和77.78%,特异度分别为80.85%、85.11%和93.62%;联合检测可提高CIN2+病变诊断的特异度。结论PAX1、TP63基因启动子甲基化水平与HPV感染阳性患者的宫颈病变关系密切,可作为临床诊断CIN2+病变的潜在辅助指标。Objective To investigate the relationship between the degree of cervical lesions and the status of paired box-1(PAX1)and tumor protein 63(TP63)gene promoter methylation in human papillomavirus(HPV)-positive patients with cervical lesions,as well as analyze their clinical significance.Methods Cervical tissue specimens were collected from 128 patients who were suspected of cervical lesions and HPV infection,and admitted to Qionghai People's Hospital between December 2021 and December 2022.According to pathological examination results,the patients were divided into the lowgrade squamous intraepithelial lesion(LSIL)group(n=43),high-grade squamous intraepithelial lesion(HSIL)group(n=51)and cervical cancer group(n=34).The second-generation hybrid capture method was used for viral load.The degree of PAX1 and TP63 gene promoter methylation in each group was detected by bisulfite sequencing,and mRNA expression of PAX1 and TP63 was detected by real-time quantitative PCR.The diagnostic performance of the degree of PAX1 and TP63 methylation for cervical intraepithelial neoplasia(CIN2+)was evaluated.Results There were statistically significant differences in HR-HPV viral load between the groups(P>0.05).A total of 49(38.28%)patients with PAX1 gene promoter methylation,and 55(42.97%)patients with TP63 gene promoter methylation were detected among the 128 patients.The percentages of PAX1 and TP63 gene promoter methylation in the cervical cancer group,HSIL group and LSIL group were(67.65%and 73.53%),(43.14%and 49.02%)and(9.30%and 11.63%),with statistically significant differences between groups(P<0.05).The mRNA expression levels of PAX1 and TP63 in the cervical cancer group,HSIL group and LSIL group were[(0.34±0.08)and(0.45±0.13)],[(0.72±0.11)and(0.63±0.09)],[(1.04±0.09)and(0.87±0.11)],with statistically significant differences between groups(P<0.05).Receiver operating characteristic(ROC)curve analysis showed that the area under the curve(AUC)values of PAX1,TP63 gene promoter methylation,and their combination for diagnosing

关 键 词：人乳头瘤状病毒 宫颈癌 盒家族基因1 肿瘤蛋白63 甲基化 

分 类 号：R511[医药卫生—内科学]