Background <p>Cytotoxic drug (CD) hazard is a significant concern in healthcare provision, affecting nurses through various exposure means. It is important to limit the risks of exposure by enhancing nurses’ knowledge of CD and improving compliance with evidence-based practices.</p> Methods <p>This study aimed to assess the knowledge, practice, and attitude of nurses working in an oncology specialty hospital by implementing an educational program. This is a pre-post test quasi-experimental study design without a control group. We utilized a simple random sampling approach to select 160 nurses. The research team compared the results before and after the intervention.</p> Results <p>Most of the nurses acquired their knowledge on the use of antineoplastic agents through organizational mandatory training programs. While knowledge improved significantly post-intervention (<i>p</i> = 0.002), changes in self-reported practice (<i>p</i> = 0.11) and attitude (<i>p</i> = 0.22) were not statistically significant. This suggests that a single educational session is sufficient to improve knowledge but insufficient to change behavior or attitudes in the short term. All three knowledge, practice, and attitude scores were positively and significantly correlated with years of work experience and the age of nurses. Both academic education and training programs have a significant impact on the knowledge and behavior of nurses.</p> Conclusion <p>Nurses demonstrated greater awareness after the education intervention. Nursing continuous education and safety professionals are advised to adopt such educational interventions within a holistic organizational behavioral change program to impact long-term nurses’ behavior.</p>