探讨人乳头瘤病毒(HPV) E6/E7 mRNA检测在宫颈上皮内瘤变(CIN)1患者预后中的预测价值。

选取2013年7月至2014年10月，于烟台市烟台山医院妇科经阴道镜下宫颈活组织病理学检查，首次确诊为CIN1的107例患者为研究对象。所有受试者均于首次确诊为CIN1前1个月内，同时接受宫颈液基薄层细胞学检查(TCT)、HPV DNA基因分型及HPV E6/E7 mRNA检测；于阴道镜下宫颈活组织病理学检查确诊为CIN1后，对其进行为期2年的随访，每6个月随访1次。随访检查项目包括宫颈TCT、HPV DNA基因分型及HPV E6/E7 mRNA检测，并对可疑病变者予以阴道镜下宫颈活组织病理学检查。根据随访结束时受试者的TCT、HPV DNA基因分型、HPV E6/E7 mRNA检测结果及阴道镜下宫颈活组织病理学检查结果，将本研究受试者分为转阴组与持续或进展组。采用χ²检验，对2组受试者的HPV E6/E7 mRNA阳性率进行比较；采用Mann-Whitney U秩和检验，对2组HPV E6/E7 mRNA表达水平进行比较；计算3种宫颈病变筛查方法对于预测CIN1预后的敏感度和特异度；绘制HPV E6/E7 mRNA表达水平预测CIN1预后的受试者工作特征(ROC)曲线，计算ROC曲线下面积(ROA-AUC)，根据约登指数最大原则，确定HPV E6/E7 mRNA表达水平预测CIN1预后的最佳临界值，同时计算其敏感度和特异度。本研究遵循的程序符合烟台市烟台山医院伦理委员会所制定的标准，获得该伦理委员会批准，并与受试者均签署临床研究知情同意书。

①根据随访结束时受试者的TCT、HPV DNA基因分型、HPV E6/E7 mRNA检测结果及阴道镜下宫颈活组织病理学检查结果，将本研究受试者分为转阴组(n＝63，随访结束时，TCT、HPV DNA基因分型及HPV E6/E7 mRNA检测结果均为正常，或阴道镜下宫颈活组织病理学检查结果为阴性)，持续或进展组(n＝44，随访结束时，阴道镜下宫颈活组织病理学检查结果为CIN1及以上)。2组患者的年龄等一般临床资料比较，差异均无统计学意义(P>0.05)。②持续或进展组受试者确诊为CIN1时的HPV E6/E7 mRNA阳性率及其中位表达水平分别为61.9%(39/63)和3 738.4 copy/mL，转阴组分别为81.8%(36/44)和583.5 copy/mL，持续或进展组HPV E6/E7 mRNA阳性率及其中位表达水平均显著高于转阴组，并且差异均有统计学意义(χ²＝4.901，P＝0.027；U＝821.000，P<0.001)。③HPV-16/18阳性患者CIN持续或进展率为50.8%(31/61)，显著高于HPV-16/18阴性者的28.3%(13/46)，并且差异有统计学意义(χ²＝5.512，P＝0.019)。HPV E6/E7 mRNA阳性患者CIN持续或进展率为48.0%(36/75)，显著高于HPV E6/E7 mRNA阴性者的25.0%(8/32)，并且差异亦有统计学意义(χ²＝4.901，P＝0.027)。TCT、HPV DNA基因分型及HPV E6/E7 mRNA定性检测对于预测CIN1患者预后的敏感度分别为50.0%、70.5%、81.8%，特异度分别为66.7%、52.4%、38.1%。④HPV E6/E7 mRNA表达水平预测CIN1患者预后的ROC曲线分析结果显示，HPV E6/E7 mRNA表达水平预测CIN1持续或进展的ROC-AUC为0.704(95%CI：0.601～0.806，P<0.001)，HPV E6/E7 mRNA表达水平预测CIN持续或进展的最佳临界值为2 724.0 copy/mL，此时其预测CIN1持续或进展的敏感度为54.5%，特异度为81.0%。

HPV E6/E7 mRNA定性检测结果对于预测CIN1患者预后的敏感度较高，而其定量检测结果对于预测CIN1患者预后的特异度较高。HPV E6/E7 mRNA检测可能对CIN1患者预后的预测具有一定临床价值。

To evaluate the predictive values of human papilloma virus (HPV) E6/E7 mRNA test in the prognosis of patients with cervical intraepithelial neoplasia (CIN) 1.

From July 2013 to October 2014, a total of 107 cases of patients who were initially diagnosed as CIN1 by pathologic results of colposcopic cervical biopsy in Department of Gynecology, Yantaishan Hospital of Yantai were selected as research subjects. All subjects received cervical ThinPrep liquid-based cytology test (TCT), HPV DNA genotyping test and HPV E6/E7 mRNA test within one month prior to initial diagnosis of CIN1. Two-year follow-ups and once every six months were conducted on all subjects, including the re-examination of TCT, HPV DNA genotyping test and HPV E6/E7 mRNA test. And cervical biopsies were conducted on suspicious lesions. According to the results of TCT, HPV DNA genotyping test, HPV E6/E7 mRNA test and colposcopic cervical biopsy at the end of follow-up, all subjects were enrolled into negative group and continuous or progressive group, respectively. Chi-square test was used to compare the positive rates of HPV E6/E7 mRNA between two groups. Mann-Whitney U rank sum test was used to compare expression levels of HPV E6/E7 mRNA between two groups. The sensitivities and specificities of these three cervical screening methods in predicting the prognosis of CIN1 were calculated respectively. Then receiver operator characteristic (ROC) curve of HPV E6/E7 mRNA expression level in predicting the prognosis of CIN1 was drawn, and the area under ROC curve (ROC-AUC) was calculated.The optimal critical value of HPV E6/E7 mRNA expression level in predicting the prognosis of CIN1 was obtained when the Youden index reaching the maximum value.And its sensitivity and specificity were calculated. This study was approved by the Ethics Committee of Human Beings in Yantaishan Hospital of Yantai and informed consent of clinical research has been signed with every subject.

①According to the results of TCT, HPV DNA genotyping test, HPV E6/E7 mRNA test and colposcopic cervical biopsy at the end of follow-up, all subjects were enrolled into negative group (n=63, the results of TCT, HPV DNA genotyping test and HPV E6/E7 mRNA test all were normal or the colposcopic cervical biopsy result was negative at the end of follow-up) and continuous or progressive group (n=44, the colposcopic cervical biopsy result was CIN1 or higher grade at the end of follow-up). There were no statistically significant differences between two groups in basic clinical data such as age and etc. (P>0.05). ②The positive rate of HPV E6/E7 mRNA and median expression level of HPV E6/E7 mRNA were 61.9% (39/63) and 3 738.4 copy/mL in continuous or progressive group, which were significantly higher than those in negative group (81.8%, 36/44) and 583.5 copy/mL, respectively), and both the differences were statistically significant (χ²=4.901, P=0.027; U=821.000, P<0.001). ③According to the results of two-year follow-up, the continuous or progressive rate of CIN in patients with HPV-16/18 positive was 50.8% (31/61), which was significantly higher than that in patients with HPV-16/18 negative (28.3%, 13/46), and the difference was statistically significant (χ²=5.512, P=0.019). The continuous or progressive rate of CIN in patients with HPV E6/E7 mRNA positive was 48.0% (36/75), which was significantly higher than that in patients with HPV E6/E7 mRNA negative (25.0%, 8/32), and the difference was statistically significant (χ²=4.901, P=0.027). The sensitivities of TCT, HPV DNA genotyping test and HPV E6/E7 mRNA qualitative test in predicting the prognosis of CIN1 were 50.0%, 70.5%, 81.8%, and the specificities were 66.7%, 52.4%, 38.1%, respectively. ④The results of ROC curve analysis of HPV E6/E7 mRNA expression level in predicting the prognosis of CIN1 showed that the ROC-AUC was 0.704 (95%CI: 0.601-0.806, P<0.001), and the optimal critical of value HPV E6/E7 mRNA expression level in predicting the prognosis of CIN1 was 2 724.0 copy/mL, and the sensitivity of HPV E6/E7 mRNA expression level in predicting continuous or progression of CIN was 54.5%, and the sensitivity was 81.0%.

The sensitive of HPV E6/E7 mRNA qualitative test in predicting the prognosis of CIN1 is relatively high, while the specificity of HPV E6/E7 mRNA quantitative test in predicting the prognosis of CIN1 is relatively high. HPV E6/E7 mRNA test may has clinical values in predicting the prognosis of patients with CIN1.