宫颈病变患者人乳头状瘤病毒16的生物状态和P16蛋白的表达

Biological status and P16 protein expression of Human Papilloma Virus 16 in patients with cervical lesion

目的:探讨人乳头状瘤病毒(HPV)16的存在状态及P16表达与宫颈病变程度的关系。方法:收集216例仅HPV16亚型阳性的宫颈脱落细胞学标本,按细胞学诊断结果分为未见上皮内病变细胞或恶性细胞(NILM)40例,非典型鳞状细胞-诊断意义不明(ASCUS)53例,非典型鳞状细胞-不排除上皮内高度病变(ASC-H)35例,低度鳞状上皮内病变(LSIL)32例,高度鳞状上皮内病变(HSIL)40例和鳞状细胞癌(SCC)16例。HPV的分型检测采用PCR-反向点杂交法,采用荧光定量PCR检测HPV16的E2和E6基因,根据E2/E6比值判定HPV16 DNA的存在状态,采用免疫组化法检测P16表达。结果:①确定0.77为荧光定量PCR区分游离型和混合型HPV16的界值。②HPV16整合发生率与不同程度的宫颈病变有关,并且随着宫颈病变级别的升高,整合发生率呈上升趋势。③宫颈脱落细胞P16表达阳性率与病变程度及HPV16整合发生率呈正相关(γ=0.938,0.956 P<0.05)。结论:HPV16整合型及P16阳性表达预示宫颈病变程度加重。

Objective：To evaluate the relationship of biological status of Human Papilloma Virus（HPV） 16,expression of P16 and the degree of cervical tumorigenesis.Methods： 216 cervical exfoliative cytology specimens with only HPV16 positive were enrolled and divided into six groups according to cervical exfoliative cytology,including NILM（n=40）,ASCUS（n=53）,ASC-H（n=35）,LSIL（n=32）,HSIL（n=40） and SCC（n=16）.HPV type were detected by PCR and reverse dot blotting.Real time PCR was employed to quantify the copy number of E2 and E6 genes and analyze the biological status of HPV16 DNA according to E2/E6 and P16 protein expression was detected by immunohistochemical method.Results： The cut off value of E2/E6,used to distinguish the pure episomal form from a mixed form of episomal and integrated HPV16 DNA,was 0.77.The rate of HPV16 integration was associated with the degree of cervical lesions and the rate increased with the progression of cervical disease,and there were significant difference.A positive relation between P16 and the grade of cervical lesions,the rate of HPV16 integration was observed by Spearman analysis（γ=0.938,0.956 P〈0.05）.Conclusion： The integral mode of HPV16 and high expression of P16 protein may have predictive value for the severity of cervical lesions.