外阴鳞状上皮内瘤变皮损中人乳头瘤病毒16、18癌基因整合性研究

Integration of HPV 16/18 DNA in Vulvar Intraepithelial Neoplasia

目的探讨人乳头瘤病毒(HPV)16、18在外阴鳞状上皮内瘤变(VIN)皮损中的感染和其癌基因在人基因组中的整合情况。方法采用捕获杂交法及PCR技术筛选高危型HPV和HPV16、18DNA阳性VIN标本：经RT-PCR、巢式PCR及DNA杂交研究HPV16、18 DNA阳性VIN标本中的HPV基因转录。结果 32例VIN患者皮损中24例(75%)为高危型HPV阳性,23例为HPV 16阳性,1例为HPV 18阳性。23例HPV 16阳性标本中除1例VIN Ⅱ皮损无HPV转录外,15份标本为游离体型HPV 16基因转录,其余7例VIN Ⅲ标本呈现整合型HPV 16癌基因转录；1例HPV 18阳性VIN Ⅲ标本发现为整合型HPV 18癌基因转录。结论大部分VIN Ⅱ、Ⅲ标本存在HPV 16感染,HPV癌基因的整合多发生于VIN Ⅲ的皮损；推测高危型HPV癌基因在人基因组中的整合与VIN的发生及其向外阴鳞状细胞癌的发展有关。

Objective To investigate the pathogenic role of high risk HPVs and genomic integration of HPV 16/18 DNA in vulvar intraepithelial neoplasia （VIN） patients. Methods Hybrid Capture HPV DNA assay and PCR were performed to detect high-risk HPVs and HPV 16/18 positive samples. Then RT-PCR, nested PCR and Southern blotting analysis were used in HPV 16/18 positive samples for the amplification and analysis of papillomavirus oncogene transcripts. Results In 32 cases of VIN patients, 24（75%） high-risk HPVs were detected, 23 were HPV 16, anti only 1 was HPV 18. Genomic integration of HPV 16/18 was observed in 8 cases of VIN Ⅲ （7 HPV 16 and Ⅰ HPV 18）. Except Ⅰ case of VIN Ⅱ, 15 of 23 HPV-16 positive specimens displayed HPVI6 episomal transcripts, 7 displayed integrated transcripts. Conclusions HPV 16 is positive in most cases of VIN Ⅱ and VIN Ⅲ. Integration of HPV 16/18 oncogene only occurs in VIN Ⅲ. It is concluded that the integration of high-risk HPVs oncogene is related with the occurrence, and the development to vulvar cancer, of VIN.