Study on early diagnostic system for cervical cancer with using liquid-based cytology samples

1. 人乳头瘤病毒（HPV） 16 DNA可以整合到细胞的DNA中，从而破坏E2基因的表达，从而导致E6和E7病毒致癌基因的表达增加并发展为癌症。液体细胞学样品采用定量多次聚合酶链反应检测。每个细胞的HPV 16病毒载量从低级别鳞状上皮内病变（LSIL）降至癌性病变。三种不同的HPV 16整合状态的平均HPV 16 DNA拷贝数为：单独形式64.1，混合形式465.5，整合形式0.4。此外，单纯整合型HPV 16患者的平均年龄比偶发型和混合型患者的平均年龄大10岁以上。实时荧光定量pcr是宫颈癌LBC筛查中HPV定量和物理状态分析的有效方法。HPV L1衣壳蛋白与感染性病毒颗粒的产生一起表达，但其表达与p16表达的关系一直是宫颈HPV感染的替代标志物。对LBC材料进行L1衣壳蛋白和p16蛋白抗体免疫化学分析。L1衣壳蛋白在30%的LSILs和12%的HSILs中呈阳性，但在SCCs中为0%。相比之下，p16蛋白在55%的LSILs， 100%的HSILs和100%的SCCs中呈阳性。L1衣壳蛋白的表达随着LSILs向HSILs和SCCs的进展而降低，而p16蛋白在所有HSILs和SCCs中均呈阳性。Ll和p16表达之间的相关性表明，L1(-)/p16（+）病例有进展的可能，而L1(+)/p16（-）和L1(-)/p16（-）病例可能是非进展性病变或可能处于缓解期。

1. Human papillomavirus (HPV) 16 DNA can be integrated into the DNA of cells, thereby disrupting E2 geneexpression, which leads to increased expression of the E6 and E7 viral oncogenes and progression to cancer. Liquid-based cytology samples were examined by quantitativereal-time polymerase chain reaction. The HPV 16 viral load per cell decreased from a low-grade squamous intraepithelial lesion (LSIL) to a cancerous lesion. The average HPV 16 DNA copy numbers for three different HPV 16 integration statuses were 64.1 for the episomal form, 465.5 for the mixed form, and 0.4 for the integrated form. Furthermore, the mean age of patients with the pure integrated form of HPV 16 was more than 10 years older than those of patients with the episomal and mixed forms. Quantitative real-timePCR appears to be a useful method for quantitative and physical status analyses of HPV in cervical cancer screening with LBC samples.2. HPV L1 capsid protein is expressed together with the productionof infectious viral particles, but its expression and relation to p16 expression, which has been a surrogate marker for HPV infection in cervix. Immunochemical analyses using antibodies against L1 capsid protein and p16 protein were carried out on LBC materials. L1 capsid protein was positive in 30% of LSILs and 12% of HSILs, butin 0% of SCCs. In contrast, p16 protein was positive in 55% of LSILs, 100% of HSILs, and 100% of SCCs. Expression of L1 capsid protein decreased with lesion progression from LSILs to HSILs and SCCs, whereas p16 protein was positive in all HSILs and SCCs. The correlation between Ll and p16 expressions suggests that L1(-)/p16(+) cases have the potential for progression, whereas L1(+)/p16(-) andL1 (-)/p16(-) cases may be nonprogressive lesions or potentially in remission.