Chang Gung University

Areca nut is a significant risk factor for head and neck cancer (HNC), yet its molecular mechanisms, particularly miRNA-mediated regulation, remain poorly understood. This study investigates the regulatory networks underlying areca nut-induced HNC and explores therapeutic strategies through computational drug repurposing. Arecoline was used to assess its effects on invasion, migration, and cisplatin resistance in HNC cells and normal keratinocytes. Differentially expressed miRNAs and mRNAs were identified using high-throughput profiling, followed by integrative network analysis using the TCGA-HNSC dataset and multiMiR. OncoPredict was used for drug repurposing to identify therapeutic agents targeting dysregulated miRNA-mRNA networks. Arecoline exposure promoted invasion and cisplatin resistance, with more pronounced effects in normal keratinocytes, indicating a potential role in early tumorigenesis. Integrative transcriptomic analysis revealed a miRNA-mRNA regulatory network comprising 1971 oncogenes, 604 tumor suppressors, 35 oncogenic miRNA (OncomiRs), and 36 tumor suppressor miRNA (TSmiRs) regulating pathways related to cell motility and stress response. A tumor-suppressive network with miR-212-3p as a central hub and an oncogenic network modulated by miR-410 and miR-1-3p as critical hubs were identified. Drug repurposing analysis identified four potential therapeutic candidates (MK-2206, BYL-719, MG-132, and FGIN-1-27), with MK-2206 emerging as the most promising. MK-2206 effectively reversed arecoline-induced miRNA-mRNA dysregulation, mitigated malignant phenotypes, and selectively targeted HNC cells while sparing normal keratinocytes. This integrative approach elucidates areca nut-driven carcinogenesis through miRNA-mRNA interactions and highlights MK-2206 as a promising therapeutic strategy for areca nut-associated HNC.

Unhealthy alcohol use (UAU) screening facilitates identification of patients with alcohol use problems for intervention. This study evaluates the usefulness of gamma-glutamyl transferase (GGT) and sequential percentage of carbohydrate-deficient transferrin (%CDT) in community screening for UAU. In two indigenous communities (A and B), liver disease screening data including GGT, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alcohol use disorders identification test (AUDIT) were reviewed. In Community B, residents with high GGT were invited for sequential evaluation including %CDT, AUDIT, ultrasonography, and liver stiffness measurement. With AUDIT score ≥ 8 and ≥ 4 as UAU criteria for male and female genders, we determined the diagnostic performances and validities of GGT and sequential %CDT in detecting UAU. A total of 1755 residents with available GGT, AST, ALT, and AUDIT were reviewed. UAU prevalence ranged from 7.4% to 44% between communities. For sequential evaluation, 79 residents were enrolled in which 57 (72.2%) were classified as UAU. The performances of GGT, AST, ALT, and AST/ALT ratios in detecting UAU were 0.767, 0.66, 0.597, and 0.549, respectively. With a cutoff of GGT ≥ upper normal limit in detecting UAU, the sensitivity, specificity, accuracy, positive (PPV), and negative predictive value (NPV) were 62.5%, 75.5%, 72.1%, 47.4%, and 85.0%. In Community B, among residents with high GGT, %CDT had a performance of 0.684. With a cutoff of 1.72, the sensitivity, specificity, accuracy, PPV, and NPV were 59.6%, 81.8%, 65.8%, 89.5%, and 43.9%, respectively. GGT and sequential %CDT are useful for screening UAU in indigenous communities with high negative and positive predictive values, respectively.