摘要
目的:探讨HPV-16长调控区(LCR)4个E2蛋白结合位点(E2BSs)甲基化水平与宫颈病变的关系。方法:选取单一型HPV-16感染者的宫颈脱落细胞标本59例,分为3组。焦磷酸测序技术定量检测4个E2BSs序列包含的7个Cp G位点的甲基化水平。b DNA技术检测对应标本内E6/E7 mRNA表达水平。分析E2BSs位点甲基化水平与E6/E7 mRNA水平及宫颈病变程度的相关性。结果:HPV-16 E2BSs甲基化水平与宫颈病变均呈正相关(r_s=0.614,0.599,0.289,0.502,P均<0.05),其中E2BS3与宫颈病变间相关性较弱(r_s=0.289,P=0.027)。除E2BS3位点(P=0.070),E2BS 1、2及4位点甲基化水平分别在HPV-16无症状感染组、LSIL组和HSIL+组组间的差异均有统计学意义(P均<0.05)。LSIL组中,4个E2BSs位点间的甲基化水平差异有统计学意义(χ~2=17.672,P=0.001),且E2BS1位点甲基化水平高于E2BS 2、3及4位点甲基化水平(P均<0.05)。HSIL+组内,4个E2BSs位点间的甲基化水平比较,差异有统计学意义(χ~2=18.387,P=0.000),E2BS1位点甲基化水平高于E2BS3和E2BS4位点(P均<0.05),E2BS3位点甲基化水平最低(P均<0.05)。E6/E7 mRNA水平随E2BS1和2位点甲基化水平升高而升高(rs=0.480,0.467,P均<0.05)。E2BS1和E2BS2位点甲基化水平诊断HSIL和SCC病变的ROC曲线下面积(AUC)较大,AUC分别为0.791(95%CI=0.668~0.914)和0.806(95%CI=0.680~0.931),诊断敏感性均为73.9%,特异性分别为72.2%和80.6%。结论:HPV-16E2BSs基因甲基化状态与宫颈病变程度存在相关性,E2BSs甲基化状态有望作为临床上预测宫颈癌前病变及宫颈癌的早期分子指标。
Objective： To investigate the relationship between the methylation states of HPV-16 E2 binding sites（ E2 BSs） and cervical lesions in clinical specimens. Methods： Fiftynine cases of cervical cell samples were collected from patients with HPV-16 infection and were divided into three groups. Methylation status of the 7 Cp Gs in the four E2 BSs in HPV-16 long control region（ LCR） were quantitatively detected using pyrosequencing.Meanwhile,Quantivirus~HPV E6/E7 RNA 3.0 assay（ b DNA） was used to detect E6/E7 mRNA level in the corresponding specimens.Results： Spearman correlation analysis showed that E2 BSs methylation levels were positively correlated with cervical lesions（ r_s= 0.614,0.599,0.289 and 0.502,respec-tively; all P 0. 05）. Whereas,the correlation between E2 BS3 methylation and cervical lesions was the weakest（ r_s= 0.289,P = 0.027）.The difference of methylation levels of E2 BS1,2 and 4 sites in HPV-16 asymptomatic infection group,LSIL group and HSIL ＋ group were statistically significant（ all P 0. 05）. In the group of LSIL,the methylation levels of the four E2 BSs were significantly different（ χ~2= 17. 672,P = 0. 001）. In addition,E2 BS1 showed higher methylation level compared to E2 BS 2,3 and 4（ all P 0. 05）. In the HSIL ＋ group,statistically significant differences were found in the methylation levels of the four E2 BSs（ χ~2= 18. 387,P = 0.000）.E2 BS3 and 4 sites were less methylated than E2 BS1（ all P 0. 05）. Furthermore,the lowest methylation level was detected in E2 BS3（ all P0.05）.The E6/E7 mRNA level showed a statistically significant increase corresponding with the methylation levels of E2 BS1 and 2 sites（ rs= 0.480 and 0.467,respectively,all P0.05）. The methylation level of E2 BS1 and E2 BS2 had the best diagnostic value for predicting HSIL and SCC（ AUC = 0.791,95%CI = 0.668 ～ 0.914;AUC = 0.806,95% CI = 0.680 ～ 0.931）,which respectively achieved 73.9% sensitivity with 72.2% specificity and 73.9% sensitivity with 80.6% specificity.Conclusion： The methylation levels of HPV-16 E2 BSs were correlated with the severity of cervical lesions.The methylation status of E2 BSs may be used as an early molecular marker to predict cervical precancerous lesions and cervical cancer.
作者
李肖甫
樊婷婷
智艳芳
荣守华
李雅
李欢欢
Li Xiaofu;Fan Tingting;Zhi Yanfang(The Third Affiliated Hospital of Zhengzhou University ,Zhengzhou 45005)
出处
《现代妇产科进展》
CSCD
北大核心
2018年第4期259-262,267,共5页
Progress in Obstetrics and Gynecology
基金
2018河南省科技攻关项目(No:182102310141)
河南省医学科技攻关计划普通项目(No:201602136)
