Hotoboc Irina Elisaveta, Fudulu Alina, Huica Irina, Iancu Iulia Virginia, Grigore Raluca, Bertesteanu Șerban Vifor Gabriel, Bleotu Coralia, Anton Gabriela, Botezatu Anca
Department of Molecular Virology, "Stefan S. Nicolau" Institute of Virology, 285 Mihai Bravu Ave, 030304, Bucharest, Romania.
"Carol Davila" University of Medicine and Pharmacy, 37 Dionisie Lupu Str., 020021, Bucharest, Romania.
Rom J Intern Med. 2025 Mar 21;63(2):152-162. doi: 10.2478/rjim-2025-0005. eCollection 2025 Jun 1.
Infection with Epstein-Barr virus is a known risk factor for laryngeal carcinogenesis; it might influence DNA methylation acting as an epigenetic driver in this type of malignancy.
Paired laryngeal tissues (neoplastic and peri-neoplastic) harvested from 24 patients were included in the study. Eleven patients expressing latent/lytic EBV genes were considered positive. 5-mC% was determined using ELISA technique and TSGs (PDLIM4, WIF1, DAPK1) promoters' methylation percentages were quantified by qMS-PCR. DNMTs (DNMT1 and DNMT3B) expression levels were quantified in qRT-PCR.
Overall, in laryngeal neoplastic samples vs peri-neoplastic ones, lower 5mC% (p=0.004) and higher TSGs promoters hypermethylation were found (p<0.0001). Significant correlation between PDLIM4 and DAPK1 promoter methylation and 5-mC% (PDLIM4 p=0.0186; DAPK1 p=0.0259) was noted. Higher 5-mC% (p=0.0041), lower PDLIM4 gene promoter methylation (p=0.0017) and overexpression of DNMTs (DNMT1: p=0.0018, respectively DNMT3B: p=0.0017) were associated with EBV infection. Also, significant differences between EBV-positive and EBV-negative cases based on tumor stage (T) were noted for 5mC% in both T1/T2 (p=0.0364) and T3/T4 stages (p=0.0275), and for PDLIM4 promoter methylation in T1/T2 stages (p=0.0121).
Future studies are needed to more effectively illustrate the interplay between EBV infection and these epigenetic mechanisms. Notably, our study highlighted a correlation between EBV and epigenetic changes in laryngeal carcinoma.
感染爱泼斯坦-巴尔病毒是已知的喉癌发生风险因素；它可能通过作为这种恶性肿瘤的表观遗传驱动因素来影响DNA甲基化。
本研究纳入了从24例患者身上获取的配对喉组织（肿瘤组织和肿瘤周围组织）。11例表达潜伏/裂解性EBV基因的患者被视为阳性。使用ELISA技术测定5-mC%，并通过qMS-PCR对TSGs（PDLIM4、WIF1、DAPK1）启动子的甲基化百分比进行定量。在qRT-PCR中对DNMTs（DNMT1和DNMT3B）的表达水平进行定量。
总体而言，在喉肿瘤样本与肿瘤周围样本中，发现5mC%较低（p = 0.004），TSGs启动子的高甲基化程度较高（p < 0.0001）。注意到PDLIM4和DAPK1启动子甲基化与5-mC%之间存在显著相关性（PDLIM4 p = 0.0186；DAPK1 p = 0.0259）。较高的5-mC%（p = 0.0041）、较低的PDLIM4基因启动子甲基化（p = 0.0017）以及DNMTs的过表达（DNMT1：p = 0.0018，DNMT3B：p = 0.0017）与EBV感染相关。此外，在T1/T2期（p = 0.0364）和T3/T4期（p = 0.0275），基于肿瘤分期（T）的EBV阳性和EBV阴性病例之间在5mC%方面存在显著差异，在T1/T2期PDLIM4启动子甲基化方面也存在显著差异（p = 0.0121）。
需要进一步的研究来更有效地阐明EBV感染与这些表观遗传机制之间的相互作用。值得注意的是，我们的研究突出了EBV与喉癌表观遗传变化之间的相关性。
