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Despite the rising incidence, currently, there are no early detection methods for HPV-driven HNC (HPV-HNC). Cervical cancer studies suggest that HPV DNA methylation changes can be used as a biomarker to discriminate cancer patients from HPV-infected individuals. As such, this study was designed to establish a protocol to evaluate DNA methylation changes in HPV late genes and long control region (LCR) in saliva samples of HPV-HNC patients and HPV-positive controls. Higher methylation levels were detected in HPV late genes (L1 and L2) in both tumour and saliva samples of HPV-HNC patients compared with HPV-positive controls. Moreover, methylation patterns between tumours and corresponding saliva samples were observed to have a strong correlation (Passing-Bablok regression analysis; τ = 0.7483, P < 0.0001). Considering the differences between HNC and controls in methylation levels in late genes, and considering primer amplification efficiencies, 13 CpG sites located at L1 and L2 genes were selected for further evaluation. A total of 18 HNC saliva samples and 10 control saliva samples were assessed for the methylation levels in the selected sites. From the CpG sites evaluated statistically significant differences were identified for CpG sites at L2-CpG 6 (P = 0.0004), L1-CpG 3 (P = 0.0144), L1-CpG 2 (P = 0.0395) and L2-CpG 19 (P = 0.0455). Our pilot data indicate that higher levels of DNA methylation in HPV late genes are indicative of HPV-HNC risk, and it is a potential supplementary biomarker for salivary HPV detection-based HPV-HNC screening., (© The Author(s) 2024. Published by Oxford University Press.)

Oral cancer (OC) is the most common form of head and neck cancer. Despite the high incidence and unfavourable patient outcomes, currently, there are no biomarkers for the early detection of OC. This study aims to discover, develop, and validate a novel saliva-based microRNA signature for early diagnosis and prediction of OC risk in oral potentially malignant disorders (OPMD). The Cancer Genome Atlas (TCGA) miRNA sequencing data and small RNA sequencing data of saliva samples were used to discover differentially expressed miRNAs. Identified miRNAs were validated in saliva samples of OC (n = 50), OPMD (n = 52), and controls (n = 60) using quantitative real-time PCR. Eight differentially expressed miRNAs (miR-7-5p, miR-10b-5p, miR-182-5p, miR-215-5p, miR-431-5p, miR-486-3p, miR-3614-5p, and miR-4707-3p) were identified in the discovery phase and were validated. The efficiency of our eight-miRNA signature to discriminate OC and controls was: area under curve (AUC): 0.954, sensitivity: 86%, specificity: 90%, positive predictive value (PPV): 87.8% and negative predictive value (NPV): 88.5% whereas between OC and OPMD was: AUC: 0.911, sensitivity: 90%, specificity: 82.7%, PPV: 74.2% and NPV: 89.6%. We have developed a risk probability score to predict the presence or risk of OC in OPMD patients. We established a salivary miRNA signature that can aid in diagnosing and predicting OC, revolutionising the management of patients with OPMD. Together, our results shed new light on the management of OC by salivary miRNAs to the clinical utility of using miRNAs derived from saliva samples., (© 2024. The Author(s).)

Background: Immunocytochemistry involving single or double antibodies can be used to support or confirm a diagnosis in difficult cytology cases. There are a few reports on the application of a cocktail of 3 antibodies in the literature. We would like to report our experience using this technique in a difficult case of cervical lymph node fine-needle aspiration cytology (FNAC)., Case: A 75-year-old woman presented with a left neck mass for which FNAC was performed. The aspirates were cellular comprising atypical cells present in loose aggregates. The cells exhibited an ovoid and spindled morphology with no evidence of keratinization or any other differentiating features. The cytology was suggestive of metastatic poorly differentiated neoplasm; however, the presence of ovoid histiocytic-looking cells required further assessment. Triple immunostaining using a cocktail of CK5/6, CK34, and CD68 was performed. The spindled atypical cells were positive for CK5/6 and CK34 (brown cytoplasmic staining) and the ovoid histiocytic-looking cells were positive for CD68 (red cytoplasmic staining). Based on immunostaining, the cytological diagnosis of metastatic squamous cell carcinoma was confirmed., Conclusion: In view of our findings, the utility of triple immunocytochemical staining in cytology smears of poorly differentiated malignancy and a brief review of the literature are highlighted in this report., (Copyright © 2011 S. Karger AG, Basel.)