Prostate cancer continues to represent a significant cause of morbidity and mortality on a global scale, with tumour angiogenesis identified as a crucial factor in its progression. Vascular endothelial growth factor A (VEGF-A), a critical regulator of angiogenesis, is increasingly being acknowledged as a potential factor influencing cancer severity and patient outcomes. The present study performed a novel and comprehensive analysis of VEGF-A gene mutations in patients diagnosed with prostate cancer, exploring their prevalence, clinical associations and potential as therapeutic targets. DNA samples from 50 patients were subjected to analysis using Sanger sequencing, the results of which revealed notable findings. The present study identified that 64% of the cohort exhibited one or more VEGF-A mutations, with c.346C > G (18%) and c.380G > C (16%) representing the most prevalent missense mutations. It is noteworthy that frameshift mutations, including c.330delA (16%) and c.338delA (6%), were also identified. These genetic variations were found to be significantly associated with more aggressive cancer phenotypes. In particular, individuals with the mutation exhibited significantly higher Gleason scores, with 28% presenting scores of ≥ 7, compared to none among those without the mutation ( P = 0.001). On the whole, the findings of the present study indicate that VEGF-A mutations have the potential to serve as significant prognostic biomarkers and therapeutic targets, thereby providing crucial insight into the development of personalised treatment strategies for prostate cancer. By identifying novel mutations and their association with tumour aggressiveness, these findings contribute valuable knowledge to global prostate cancer research and may facilitate the development of targeted therapies.