Establishment and Characterization of a New Immortalized Human Oral Keratinocyte (HOK) Cell Line Harboring Various TP53 Mutations

Head and neck squamous cell carcinoma (HNSCC) is characterized by frequent TP53 mutations, which include gain-of-function (GOF) variants that drive tumor progression and chemoresistance. While genomic evidence across multiple invasive and metastatic specimens indicates TP53 loss or mutation occurs relatively early in HNSCC carcinogenesis, it has been difficult to discern how TP53 mutation drives further tumor evolution and influences the tumor microenvironment. To address this question, we generated an immortalized human oral keratinocyte (HOK) cell line expressing various TP53 mutant forms to determine how mutations impact functional and genomic characteristics of HOKs that can in turn drive their evolution to neoplastic cells. Key results revealed significant phenotypic and molecular divergence: high-risk lines exhibited enhanced invasiveness and chemoresistance compared to low-risk counterparts. Weighted gene co-expression network analysis (WGCNA) linked high-risk mutations to pro-metastatic pathways and stress-response signatures, with the C238F line uniquely enriched for p53-related GOF mechanisms.