Department of Molecular Bases of Medicine, 1st Chair of Internal Diseases, Medical University of Lodz, Pomorska 251, C5, 92-213, Lodz, Poland.
Med Oncol. 2014 Mar;31(3):842. doi: 10.1007/s12032-014-0842-6. Epub 2014 Jan 17.
Integration of the HPV genome into a host cell DNA leads to the deregulated overexpression of the viral E6 and E7 oncoproteins, and this is a key factor for progression from low-grade cervical lesions to high-grade lesions and invasive cervical cancer. The aim of our study was to analyze the expression levels of HPV E6I/E6II and E7 genes in cervical neoplasia of different grades. The analysis involved 10 low-grade squamous intraepithelial lesions (CIN1), 15 high-grade lesions (CIN2 and CIN3), as well as normal cytology samples (n=10). HPV genotyping was done using RealLine HPV 16/18 kit. The expression analysis was performed in real-time PCR assay using gene-specific primers and SYBR Green. HPV16 DNA was found in 65.71% patients, including also normal cytology samples. The increased expression level of E6I was observed in 12 (34.3%) patients. The expression of E6II was increased in 10 (28.6%) samples, and E7 overexpression was found in 14 (40%) patients. Significant positive correlation was observed between the amount of HPV16 DNA and the levels of E6I and E6II expression. There were no statistically significant differences in expression levels of the studied genes between the groups (CIN1 vs. CIN2/CIN3 vs. normal cytology). Statistically significant differences were found in CIN2/CIN3 group, with the higher expression of E6II as compared with E6I. We suggest that the expression level of E6*II gene might be used as an indicator of cervical cancer severity, in patients with high-grade cervical neoplasia, but these observations need to be confirmed in a larger patient cohort.
HPV 基因组整合到宿主细胞 DNA 中会导致病毒 E6 和 E7 癌蛋白的失调过表达，这是低级别宫颈病变进展为高级别病变和浸润性宫颈癌的关键因素。我们的研究目的是分析不同级别宫颈病变中 HPV E6I/E6II 和 E7 基因的表达水平。该分析涉及 10 例低级别鳞状上皮内病变（CIN1）、15 例高级别病变（CIN2 和 CIN3）以及正常细胞学样本（n=10）。使用 RealLine HPV 16/18 试剂盒进行 HPV 基因分型。使用基因特异性引物和 SYBR Green 在实时 PCR 测定中进行表达分析。在 65.71%的患者中发现了 HPV16 DNA，包括正常细胞学样本。在 12 名（34.3%）患者中观察到 E6I 的表达水平增加。在 10 名（28.6%）样本中观察到 E6II 的表达增加，14 名（40%）患者中发现 E7 过表达。HPV16 DNA 量与 E6I 和 E6II 表达水平之间存在显著正相关。在研究基因的表达水平在 CIN1 组、CIN2/CIN3 组和正常细胞学组之间没有统计学差异。在 CIN2/CIN3 组中发现了统计学上的显著差异，E6II 的表达高于 E6I。我们认为，在患有高级别宫颈肿瘤的患者中，E6*II 基因的表达水平可能可用作宫颈癌严重程度的指标，但这些观察结果需要在更大的患者队列中得到证实。
