Glioblastoma Multiforme (GBM) is one of the most lethal forms of malignant primary brain tumors. To date, untreated GBM can be fatal within 3 months or 12 months if treatment follows initial onset. Novel and innovative treatments are crucial. Cancer treatments that involve targeted therapy include DNA intercalators, shown to stack between DNA bases and affect cell death in abnormal cell growth. Known DNA intercalators, include antineoplastic ellipticine and imaging agent ethidium bromide (EtBr). A recent GBM clinical trial agent, Berberine (BBR), remains unclear in anticancer-activity exact mechanism and efficacy. Our evidence-based studies revealed BBR chemical structure supports its investigation as a candidate DNA intercalator. Our research goal is to discover proof-of-concept for BBR anti-neoplastic mechanism of action as a DNA intercalator. A computational model of BBR stacked between DNA base pairs generated frontier molecular orbital energies and interaction energies via ab initio calculations. Verification of these energies with those known for ellipticine stacked between DNA base pairs suggests BBR mechanism of action uses DNA intercalation. These preliminary results can lead to berberine redesign or combined treatment strategies. Future BBR characterization studies will capture expanded computational models and empirical studies: structure-activity-relationships, kinetic binding, preclinical ex vivo and in vitro/ vivo cancer models.