Gashi G, Mahovlić V, Bahtiri E, Kurshumliu F, Podrimaj-Bytyqi A, Elezaj I R
a Department of Pathology, Faculty of Medicine , University of Prishtina , Prishtina , Kosovo.
b Department of Pathology and Cytology, Clinical Unit of Gynecological Cytology , University Hospital Center , Zagreb , Croatia.
Biotech Histochem. 2018;93(7):496-504. doi: 10.1080/10520295.2018.1462533. Epub 2018 May 23.
Human papilloma virus (HPV) infection is the main etiological factor for cervical intraepithelial lesions (CIN). An important characteristic of this process is the loss of genome stability. Therefore, it is imperative to use biomarkers of DNA damage caused by genomic instability to identify high risk individuals. We investigated the frequency of micronuclei (MN) in peripheral blood lymphocytes (PBL) of 20 patients, diagnosed as histologically CIN 1 and 10 healthy controls. We also examined the frequency of other nuclear anomalies including nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) in PBL of patients with CIN 1 and healthy controls, and evaluated the benefits of p16 and Ki-67 (p16/Ki-67) immunohistochemical double staining for identifying cervical squamous cells that express HPV E6/E7 oncogenes. We analyzed the association between the frequency of MN in PBL and the amount of p16/Ki-67 co-expression in CIN 1 patients to establish genomic instability. Among CIN 1 subjects, 15% exhibited diffuse p16/Ki-67 co-expression and were considered high positive, 25% of the CIN 1 cases exhibited p16/Ki-67 co-expression restricted to the lower part of the epithelium and were considered low positive and the remaining 60% of cases were negative. The frequency of MN, NPBs and NBUDs differed significantly among groups. We found a statistically significant positive correlation between p16/Ki-67 co-expression and the frequency of MN, NPBs and NBUDs in PBL. Our findings demonstrate the efficacy of p16/Ki-67 double immunostaining for histological samples with CIN 1. MN frequency in PBL might be useful for detecting genomic instability in cases of HPV infection and CIN.
人乳头瘤病毒（HPV）感染是宫颈上皮内瘤变（CIN）的主要病因。这一过程的一个重要特征是基因组稳定性的丧失。因此，利用基因组不稳定导致的DNA损伤生物标志物来识别高危个体势在必行。我们调查了20例经组织学诊断为CIN 1的患者和10名健康对照者外周血淋巴细胞（PBL）中的微核（MN）频率。我们还检查了CIN 1患者和健康对照者PBL中其他核异常的频率，包括核质桥（NPB）和核芽（NBUD），并评估了p16和Ki-67（p16/Ki-67）免疫组化双重染色对识别表达HPV E6/E7癌基因的宫颈鳞状细胞的益处。我们分析了CIN 1患者PBL中MN频率与p16/Ki-67共表达量之间的关联，以确定基因组不稳定性。在CIN 1受试者中，15%表现为弥漫性p16/Ki-67共表达，被认为是高阳性，25%的CIN 1病例表现为p16/Ki-67共表达局限于上皮下部，被认为是低阳性，其余60%的病例为阴性。各组之间MN、NPB和NBUD的频率差异显著。我们发现p16/Ki-67共表达与PBL中MN、NPB和NBUD的频率之间存在统计学上显著的正相关。我们的研究结果证明了p16/Ki-67双重免疫染色对CIN 1组织学样本的有效性。PBL中的MN频率可能有助于检测HPV感染和CIN病例中的基因组不稳定性。
