Szostek Sława, Zawilińska Barbara, Klimek Małgorzata, Wójcik Kinga, Koprynia Małgorzta, Kosz-Vnenchak Magdalena
Zakład Wirusologii, Katedra Mikrobiologii, Uniwersytet Jagielloński Collegium Medicum w Krakowie, Polska.
Ginekol Pol. 2011 Jun;82(6):441-5.
Persistent high-risk HPV infection, especially HPV-16, is considered to be an important step in the process of cervical carcinogenesis. Integration of viral DNA into the host genome through the destruction of HPV E2 sequences, increases the expression of viral proteins E6 and E7 and their participation in the transformation of cervical cancer.
The aim of this study was to apply real-time PCR (RT-PCR) to assess the prevalence of integrated and episomal HPV-16 DNA and determine viral DNA load in women with cervical intraepithelial lesions and invasive cervical cancer
A total of 84 women infected with HPV-16, including 44 with LSIL, 7 with HSIL and 33 with invasive cervical cancer participated in the study Cervical specimens were collected using the cytobrush. The presence of a sequence of E2 and E6 HPV-16 and human gene RNasy P was detected by quantitative RT-PCR. The viral load presented as the form of the virus genome copy numbers per 1,000 cells.
The integrated form of HPV-16 genome was found in 97% of women with cervical cancer. In women with LSIL and HSIL mixed form (simultaneous occurrence of an integrated and episomal form) of the viral genome (84% and 57%, respectively) prevailed. The frequency of the integrated HPV-16 DNA increased with progression of dysplastic lesions of the cervix (p<0.001). Statistically significant differences in average number of copies of the virus in women with LSIL and HSIL compared to patients with cancer (p<0.001) were observed. The highest viral load was detected in women demonstrating an integrated HPV-16 DNA.
Quantitative analysis of the sequence of E2 and E6 HPV-16 tested by RT-PCR can be used to determine the degree of integration of the viral genome and quantitative evaluation of viral load in clinical material. It can also serve as an additional parameter defining risk of progression of transformation in the cervix.
持续性高危型人乳头瘤病毒（HPV）感染，尤其是HPV-16感染，被认为是子宫颈癌发生过程中的重要一步。病毒DNA通过破坏HPV E2序列整合到宿主基因组中，增加了病毒蛋白E6和E7的表达及其在子宫颈癌转化中的参与。
本研究旨在应用实时荧光定量聚合酶链反应（RT-PCR）评估整合型和游离型HPV-16 DNA的流行情况，并确定子宫颈上皮内瘤变和浸润性子宫颈癌女性的病毒DNA载量。
共有84例HPV-16感染女性参与研究，其中44例为低度鳞状上皮内病变（LSIL），7例为高度鳞状上皮内病变（HSIL），33例为浸润性子宫颈癌。使用细胞刷收集子宫颈标本。通过定量RT-PCR检测HPV-16 E2和E6序列以及人类基因RNasy P的存在情况。病毒载量以每1000个细胞中病毒基因组拷贝数的形式呈现。
97%的子宫颈癌女性中发现了HPV-16基因组的整合形式。在LSIL和HSIL女性中，病毒基因组的混合形式（整合型和游离型同时出现）占主导（分别为84%和57%）。整合型HPV-16 DNA的频率随着子宫颈发育异常病变的进展而增加（p<0.001）。与癌症患者相比，LSIL和HSIL女性的病毒平均拷贝数存在统计学显著差异（p<0.001）。在显示整合型HPV-16 DNA的女性中检测到最高病毒载量。
通过RT-PCR检测HPV-16 E2和E6序列的定量分析可用于确定病毒基因组的整合程度，并对临床材料中的病毒载量进行定量评估。它还可作为定义子宫颈转化进展风险的一个附加参数。
