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目的通过检测不同宫颈病变组织中Foxp3+调节性T细胞(regulatory T cell,Treg)的表达及人乳头瘤病毒(human papilloma virus,HPV)基因组整合状态,研究Treg与人乳头瘤病毒整合状态在宫颈病变发病机制中的作用及意义。方法选取2012年11月至2013年7月间,中国医科大学附属盛京医院妇产科住院的119例单一HPV58阳性患者的宫颈脱落细胞标本及相应宫颈组织石蜡标本。包括正常宫颈22例、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)79例、宫颈癌(cervical cancer,CC)18例。多重PCR技术同时检测HPV58型E2和E6基因,通过E2/E6比值法评估HPV58型基因组在不同宫颈病变中的整合状态。免疫组化方法检测宫颈组织中Foxp3表达情况。结果 HPV58的总整合率为81.51%,HPV58的整合率在正常组中为72.73%,CINⅠ组为76.19%,CINⅡ组为81.82%,CINⅢ组为88.00%,CC组为88.89%。HPV58的整合率随宫颈病变加重呈逐渐升高,差异有统计学意义(χ2=8.399,P=0.004,线性趋势χ2检验)。Foxp3表达强度与宫颈病变严重程度呈正相关,差异有统计学意义(r s=0.538,P=0.000,Spearman等级相关分析)。Foxp3表达强度在HPV不同整合状态下是不同的,差异有统计学意义(χ2=12.058,P=0.002,Kruskal Wallis检验)。结论 HPV58基因组的整合与Foxp3+Treg在宫颈病变发展中密切相关。
Objective To investigate the relationship between Treg and human papillomavirus (HPV) integration in cervical lesions by detecting the expression of Foxp3 + regulatory T cell (Treg) and the integrated status of human papilloma virus (HPV) genome in different cervical lesions. The role of pathogenesis and its significance. Methods From November 2012 to July 2013, specimens of cervical exfoliated cells and corresponding cervical tissue from 119 single HPV58-positive patients hospitalized in Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University were selected. Including 22 cases of normal cervix, 79 cases of cervical intraepithelial neoplasia (CIN) and 18 cases of cervical cancer (CC). Multiplex PCR was used to detect the E2 and E6 genes of HPV58 simultaneously, and the integration status of HPV58 genotypes in different cervical lesions was evaluated by E2 / E6 ratio method. Immunohistochemical detection of Foxp3 expression in cervical tissue. Results The overall HPV58 integration rate was 81.51%. The HPV58 integration rate was 72.73% in the normal group, 76.19% in the CINⅠ group, 81.82% in the CINⅡ group, 88.00% in the CINⅢ group and 88.89% in the CC group. HPV58 integration rate increased with the severity of cervical lesions was gradually increased, the difference was statistically significant (χ2 = 8.399, P = 0.004, linear trend χ2 test). There was a positive correlation between the expression of Foxp3 and the severity of cervical lesions (r s = 0.538, P = 0.000, Spearman rank correlation analysis). Foxp3 expression intensity in different integrated status of HPV is different, the difference was statistically significant (χ2 = 12.058, P = 0.002, Kruskal Wallis test). Conclusion The integration of HPV58 genome is closely related to the development of Foxp3 + Treg in cervical lesions.