Abstract <p>Translational medicine plays a crucial role in bridging basic research and clinical application, driving innovations in cancer diagnostics and treatment. Recent advances in cancer research have reshaped our understanding of oncological diseases and opened new directions for more precise and effective therapies. Next-generation sequencing (NGS) and third-generation sequencing (TGS) now provide detailed insights into the molecular mechanisms underlying tumor development, heterogeneity, and progression, and are increasingly adopted in both research and clinical settings. These technologies are accelerating the development of more tailored therapies and aiding in the discovery of novel biomarkers and molecular targets. This review highlights recent advances in NGS technologies and their growing impact on translational oncology. We discuss the strengths and limitations of different sequencing approaches, review bioinformatic tools developed for data processing and interpretation, and explore their integration with multi-omics and artificial intelligence–driven methods. Continued innovation and deeper clinical integration of these approaches promise to further refine precision oncology and improve patient outcomes.</p>