In this study, imatinib-sensitive K-562S cells were exposed to CBD, which inhibited cell proliferation in a dose-dependent manner, with an IC50 of 17.69 μM. Additionally, CBD modulated lipid metabolism and mitochondrial processes, pointing to broader metabolic disruption in leukemia cells. Pathway analysis confirmed CBD’s influence on zinc and copper homeostasis, fatty acid transport, apoptosis networks, and immune signaling. These findings highlight CBD’s capacity to alter transcriptional regulation, immune pathways, and apoptosis in leukemia cells, reinforcing its potential role as an anticancer agent. The authors suggest that future studies should explore CBD’s use in combination therapies and its precision applications in hematological malignancies.