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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 investigate the relationship between the presence of human papillomavirus (HPV) 16 and the expression of P16 in cervical lesions. METHODS: Totally 216 cases of HPV16 subtype positive cervical cytology specimens were collected and divided into 40 cases without intraepithelial lesion cells or malignant cells (NILM) by cytological diagnosis. Atypical squamous cells with unknown diagnostic significance (ASCUS ), And atypical squamous cells-35 cases of ASC-H, 32 cases of low-grade squamous intraepithelial lesion (LSIL), 40 cases of high-grade squamous intraepithelial lesion (HSIL) 16 cases of squamous cell carcinoma (SCC). HPV typing was detected by PCR-reverse dot blot hybridization. E2 and E6 genes of HPV16 were detected by real-time PCR. The expression of HPV16 DNA was determined by the ratio of E2 / E6, and the expression of P16 was detected by immunohistochemistry. Results: ①Determination of 0.77 is the cut-off value of fluorescent quantitative PCR for distinguishing between free type and mixed type HPV16. ② The incidence of HPV16 integration with varying degrees of cervical lesions, and as the level of cervical lesions increased, the incidence of integration is on the rise. ③ The positive rate of P16 expression in cervical exfoliated cells was positively correlated with the degree of lesion and the incidence of HPV16 integration (γ = 0.938,0.956 P <0.05). Conclusion: The positive expression of HPV16 and P16 indicates that the cervical lesions are more severe.