Anti-tumor efficacy of Habb-e-Asgandh as an adjuvant therapy in chronic myelogenous leukemia in vitro

Abstract

IntroductionChronic myeloid leukemia (CML) is a myeloproliferative hematological malignancy wherein the increased relapsed cases and lack of drug effectiveness generates a need to identify an effective treatment regimen. A unani formulation, Habb-e-Asgandh (HEA) is being used for treatment of ailments such as arthritis, however, its role in CML therapeutics has not been reported yet. Hence, anti-leukemic efficacy of HEA was assessed alone or in combination with standard chemotherapeutic, imatinib in CML cell line K562. MethodsThe anti-leukemic effect of HEA was evaluated on cell cytotoxicity by MTT assay, cell cycle analysis by propidium iodide staining, apoptotic assay by Annexin-V assay, mitochondrial membrane potential (MMP) by JC1 staining. Further, effect of drug treatment on oxidative stress and cell migration was determined by ORAC assay and transwell migration assay respectively. Lastly, changes in angiogenic marker (VEGF) and inflammatory marker (IL-6) were examined by ELISA. ResultsHEA induced cytotoxicity of leukemic cells which increased further when co-treated with imatinib. The combinatorial treatment of HEA with imatinib potentiates anti-tumor effect by inducing cell apoptosis with disruption of MMP even at low concentrations. Subsequent studies revealed that tumor killing property of HEA increased the ROS generation. Furthermore, HEA in combination with imatinib dramatically reduced cell migration along with expression of VEGF and IL-6. ConclusionThis maiden study demonstrated that HEA possesses anti-leukemic potential in vitro. This formulation potentiates the effect of imatinib even at low concentrations suggesting the utilization of HEA as a novel adjuvant therapy for the better management of CML in future.