目的：分析不同病变程度的宫颈病变患者高危型人乳头瘤病毒（HPV）DNA和高危型HPV E6/E7 mRNA表达阳性率及拷贝数的差异，评估两者对宫颈癌早期筛查的临床价值。方法收集天津市中心妇产科医院2014年因宫颈疾患行阴道镜下宫颈活检的154例患者资料和因其他良性妇科疾患行全子宫切除术的32例患者资料（对照组），154例根据宫颈活检病理结果分为宫颈上皮内瘤变低级别组（51例）、高级别组（71例）及宫颈浸润癌组（32例）。应用杂交捕获技术（HC2）HPV DNA检测和荧光定量杂交捕获技术HPV E6/E7 mRNA检测进行定量测定，并对全部患者的术后或宫颈活检石蜡标本进行E6/E7蛋白免疫组化检测。结果 HC2 HPV DNA检测和HPV E6/E7 mRNA检测显示对照组阳性率低于各级别病变组（P<0.05），高危型HPV E6/E7 mRNA检测的拷贝数随着病变级别的加重而明显增高，而HC2 HPV DNA检测的拷贝数不随病变级别的加重而改变。高级别组及宫颈浸润癌组患者中，高危型HPV E6/E7 mRNA检测拷贝数>10000的患者比例明显增多。免疫组化结果显示对照组E6/E7阳性率低于各级别病变组，高级别组、宫颈浸润癌组的E6/E7阳性率高于低级别组（均P<0.05）。结论 HPV E6/E7 mRNA的表达及拷贝数的测定与宫颈病变程度密切相关，是宫颈癌早期筛查的有效措施之一，HPV DNA检测结果为阴性时，其排除疾病意义更高，而mRNA检测结果为阳性时，其预测疾病发生及进展的价值更好，两者结合使用有利于综合评价发生宫颈病变的风险。%Objective To analyze the differences of positive detection rate and copy number of human papillomavirus (HPV) DNA and E6/E7 mRNA between different grades of cervical lesions, and evaluate their clinical values in early screen⁃ing of cervical cancer. Methods The cervical exfoliated cell samples from 154 women undergoing biopsy examination and 32 objects undergoing hysterectomy (control group) were collected in Tianjin Central Hospital of Gynecology Obstetrics in 2014. According to the pathological results of cervical biopsy, 154 samples were divided into low-grade squamous intraepi⁃thelial lesion group (LSIL, n=51), high-grade squamous intraepithelial lesion group (HSIL, n=71), and squamous cell carci⁃noma group (SCC, n=32). HPV DNA was tested with hybrid capture technology, and E6/E7 mRNA was detected with fluores⁃cence quantitative hybridization. Immunohistochemistry was performed by detecting E6/E7 protein in all patients after sur⁃gery or cervical biopsy. Results Combined results of HPV DNA and E6/E7 mRNA demonstrated that the positive detection rate was significantly lower in control group than that of all levels of lesion groups (P<0.05). The copy number of high risk HPV E6/E7 mRNA was significantly increased with the aggravation of lesions (P<0.05), whereas no difference was found in that of HPV DNA. Compared with the normal control and low-grade squamous intraepithelial lesion group, cervical cancer patients with mRNA copies > 10 000 E6/E7 were significantly increased in high-grade squamous intraepithelial lesion group. Immunohistochemical results showed that the positive detection rate of E6/E7 was significantly lower in control group than that of all levels of lesion groups (P<0.05). The positive rate of E6/E7 was significantly higher in the high-grade squa⁃mous intraepithelial lesion group than that of low-grade group (P<0.05). Conclusion HPV infection is closely related to cervical abnormalities, which is one of effective measures for early screening of cervical cancer. The negative result of HPV DNA is very helpful to exclude the cervical abnormality, whereas the positive detection of mRNA has great value in predict⁃ing the disease. Combined results of positive detection and copy number make a comprehensive evaluation for the risk of cer⁃vical lesions.
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