Targeted Hotspot Mutation Analysis of KRAS, TP53, and PTEN in Endometrial Cancer Revealed by Next-Generation Sequencing

Endometrial cancer (EC) is a heterogeneous malignancy characterized by diverse molecular and clinicopathological features. This study investigated somatic mutations in three hotspot-region genes commonly implicated in EC pathogenesis (KRAS, TP53, PTEN) and their associations with clinical characteristics and biomarkers, rather than comprehensive genomic profiling. Targeted next-generation sequencing (NGS) was used to analyze hotspot regions of KRAS (exons 2 and 4), TP53 (exons 4 and 7), and PTEN (exon 2), while mutation parameters, including mutant allele frequency (MAF), allelic depth, and variant frequency, were assessed alongside clinicopathological data. Pathogenic KRAS missense mutations (p.G12D and p.A146V), TP53 missense (p.G245C) and stop-gain (p.W91*) variants, and a pathogenic PTEN mutation (p.D52Y) were identified, with MAF values ranging from 36.2% to 77.1%. One patient also exhibited microsatellite instability-high (MSI-H) status. Serum CA125 concentrations were significantly higher in serous carcinoma than in endometrioid carcinoma and non-malignant cases, suggesting potential prognostic rather than diagnostic value. This suggests an association between CA125 levels and histological subtype rather than diagnostic or prognostic value, as this study did not evaluate survival, recurrence, or longitudinal outcomes. The detected mutations highlight disruptions in estrogen signaling, DNA damage response, and PI3K/AKT pathway regulation, illustrating the diversity of alterations detectable within these three hotspot regions in EC. These findings provide insight into the mutational status of KRAS, TP53, and PTEN hotspot regions in endometrial cancer and support the potential use of identified genetic alterations as biomarkers for precision medicine and targeted therapeutic strategies, while indicating that CA125’s clinical utility in EC, whether diagnostic or prognostic, remains to be established through outcome-based studies. This also reinforces CA125’s limited diagnostic but possible prognostic role in EC management.