Prognostic impact of miR-34b/c DNA methylation, gene expression, and promoter polymorphism in HPV-negative oral squamous cell carcinomas

• Published in: Scientific reports Vol. 12; no. 1; p. 1296
• Main Authors: Supic, Gordana, Stefik, Debora, Ivkovic, Nemanja, Sami, Ahmad, Zeljic, Katarina, Jovic, Sasa, Kozomara, Ruzica, Vojvodic, Danilo, Stosic, Srboljub
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 25-01-2022; Nature Publishing Group; Nature Portfolio
• Subjects: 631/208; 631/337; 631/67; 692/4028; Adult; Aged; Aged, 80 and over; Alphapapillomavirus - genetics; Biomarkers, Tumor - genetics; Cancer; Case-Control Studies; Deoxyribonucleic acid; DNA; DNA methylation; DNA Methylation - genetics; Female; Follow-Up Studies; Gene Expression; Gene polymorphism; Gene regulation; Genotyping; Health risks; Humanities and Social Sciences; Humans; Male; Medical prognosis; Medical screening; MicroRNAs - genetics; Middle Aged; Mouth Neoplasms - genetics; Mouth Neoplasms - pathology; multidisciplinary; Oral cancer; Oral carcinoma; Oral squamous cell carcinoma; Papillomavirus Infections - diagnosis; Papillomavirus Infections - virology; Patients; Polymerase chain reaction; Polymorphism; Polymorphism, Single Nucleotide; Prognosis; Promoter Regions, Genetic; Risk assessment; Science; Science (multidisciplinary); Serbia - epidemiology; Squamous cell carcinoma; Squamous Cell Carcinoma of Head and Neck - genetics; Squamous Cell Carcinoma of Head and Neck - pathology; Survival; Therapeutic targets; Tumors; Serbia
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-05399-1

Micro RNAs (miRNAs) have a key role in gene expression regulation in cancer. The aim of the current study is to evaluate the prognostic value of miR-34b/c promoter hypermethylation, gene expression, and polymorphism in HPV-negative oral squamous cell carcinomas (OSCC). MiR-34b/c promoter hypermethylation and pre-miR-34b/c polymorphism rs4938723 were evaluated in tumor tissues of 148 patients, and miR-34b expression in 123 HPV-negative OSCC. For risk assessment, the control group was comprised of 175 healthy individuals. MiR-34b/c promoter hypermethylation was determined by methylation-specific PCR. Gene expression, genotyping and HPV screening was assessed by Q-PCR. The data from our hospital cohort indicated that miR-34b/c DNA methylation was associated with nodal status (p = 0.048), and predicted the shorter overall survival of HPV-negative OSCC patients (p = 0.008). Down-regulated miR-34b/c expression was associated with smoking (p = 0.047), alcohol use (p = 0.009), stage (p = 0.025), recurrences (p = 0.000), and a poor survival (p = 0.00029). Median values of miR-34b expression were significantly lower in advanced stages III/IV as opposed to stage I/II, p = 0.006, and in nodal positive vs negative patients (p = 0.045). TCGA data also indicated that tumors with stage I–III expressed significantly higher levels of miR-34b, compared to tumors with stage IV (p = 0.035), Low miR-34b/c expression was associated with poor survival in smokers (p = 0.001) and patients with tongue carcinomas (p = 0.00003), and TCGA analysis confirmed these findings although miR-34b expression and miR-34b/c methylation were not associated with survival outcome in the whole TCGA cohort. A significant negative miR-34b/c expression–methylation correlation was observed in our hospital cohort (p = 0.017) and in TCGA cohort. Pre-miR-34b/c polymorphism was not associated with oral cancer risk. Our findings indicate that miR-34b/c hypermethylation and low miR-34b expression could promote the progression and predict the poor prognosis for HPV-negative OSCC, which suggests miR-34b/c as a promising biomarker and therapeutic target for OSCC in the future.