女性尖锐湿疣和宫颈上皮内瘤变HPV16型E2、E6、E7基因突变及E6、E7抗原表达

Analysis on E2, E6, E7 Gene Mutations and Oncogenic E6,E7 Antigen Expressions of HPV 16 in Women Patients with CA and Cervical Intraepithelial Neoplasia

目的研究女性尖锐湿疣(CA)和宫颈上皮内瘤变(CIN)中人乳头瘤病毒(HPV)亚型分布,以及高危型HPV16的E2、E6、E7基因突变及致癌抗原E6、E7表达。方法从24例女性CA、11例2级CIN(CIN2)和23例3级CIN(CIN3)共58份石蜡包埋组织中提取HPV基因组DNA,用PCR-膜杂交法检测HPV亚型,DNA测序法检测HPV16型E2、E6、E7基因突变,免疫组化S-P法检测HPV16型E6、E7抗原表达。结果女性CA、CIN2和CIN3患者58份标本中共检出10个HPV亚型,高危型HPV检出率分别为20.8%、45.5%和60.9%,HPV16型检出率分别为12.5%、18.2%和34.8%。24例女性CA中高危型和低危型HPV检出率为20.8%和70.8%,宫颈CA高危型HPV检出率60.0%。HPV16型E2、E6、E7基因分别检出10、4和4个突变位点。首次在1例女性外阴CA和3例CIN3中一致检出HPV16型E2基因9个突变位点。HPV16型E6抗原在CA、CIN2和CIN3均有表达,但E7抗原仅在CIN3表达。结论女性宫颈CA和CIN多为高危型HPV感染,女性外阴CA的HPV16型E2基因突变可能与CIN3发生有一定关联。

Objective In order to analyze subtypes of human papillomavirus（HPV） in women patients with condyloma acuminatum（CA） and cervical intraepithelial neoplasia（CIN）, as well as mutations of E2, E6, E7 genes and expressions of oncogenic E6, E7 antigens of the high-risk subtype HPV16. Methods HPV genomic DNAs were extracted from 58formalin-fixed paraffin-embedded biopsied tissues from 24 women patients with CA, 11 patients with CIN grade 2（CIN2） and23 patients with CIN grade 3（CIN3）. HPV subtypes were tested with PCR-membrane in situ hybridization assay. Mutations in E2, E6 and E7 genes of high-risk HPV16 were assayed by DNA sequencing and then compared with wild type of HPV16 in Gen Bank. Oncogenic E6 and E7 antigens of HPV16 were immunohistochemically stained with a streptavidin-peroxidase（SP） method. Results A total of ten subtypes of HPV were detected in women patients with CA, CIN2 and CIN3. Among which, high-risk subtypes were detected in 20.8% of CA, 45.5% of CIN2 and 60.9% of CIN3, respectively. HPV16 was detected in 12.5% of CA, 18.2% of CIN2 and 34.8% of CIN3, respectively. Both high-risk（20.8%） and low-risk（70.8%） of HPV subtypes were detected in women patients with CA, and high-risk subtypes were detected in 60.0% of lesions on cervix.There were totally 10, 4 and 4 mutations detected in E2, E6 and E7 genes of HPV16. Nine consistent substitution mutations in E2 gene of HPV16 were detected for the first time in both vulvar lesion of a woman patient with CA and cervical lesions of three patients with CIN3. HPV16 E6 antigens were expressed in CA, CIN2 and CIN3, whereas HPV16 E7 antigens were expressed only in CIN3. Conclusion CA on cervix and CIN were most commonly infected with high-risk subtypes of HPV.Mutations in E2 gene of HPV16 in women vulva CA lesion might have a link to the pathogenesis of CIN3.