Title:Determination of the Apoptotic Effect and Molecular Docking of Benzamide Derivative XT5 in K562 Cells
VOLUME: 18 ISSUE: 11
Author(s):Tulin Ozkan*, Yalda Hekmatshoar*, Tugba Ertan-Bolelli, Andry N. Hidayat, Meral Beksac, Esin Aki-Yalcin, Ismail Yalcin and Asuman Sunguroglu
Affiliation:Department of Medical Biology, School of Medicine, Ankara University, Ankara, Department of Medical Biology, School of Medicine, Ankara University, Ankara, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Ankara University, Ankara, Biotechnology Institute, Ankara University, Ankara, Department of Hematology and Cord Blood Bank, Ankara University, Ankara, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Ankara University, Ankara, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Ankara University, Ankara, Department of Medical Biology, School of Medicine, Ankara University, Ankara
Keywords:CML, K562, imatinib resistance, benzamide, apoptosis.
Abstract:Background: The tyrosine kinase inhibitor, imatinib, used as a first line treatment in Chronic Myeloid
Leukemia (CML) patients, may lead to resistance and failure to therapy. Novel combinations of imatinib
with other drugs is a strategy to improve treatment efficiency.
Objective: In this study, the antileukemic and apoptotic effects of a benzamide derivative XT5 and benzoxazole
derivative XT2B and their combination with imatinib were investigated in imatinib-sensitive (K562S) and
imatinib-resistant (K562R) CML cells.
Methods: In vitro cytotoxicity was determined by MTT assay. Then, apoptotic effect of XT5 on CML cell lines
was tested by Annexin V flow cytometry, caspase activation and RT-PCR. Docking calculation was performed
using AutoDock Vina in PyMOL environment using AutoDock/Vina plugin for PyMOL.
Results: According to our MTT assay data, XT5 indicated significant antiproliferative effect on cell lines, therefore
we investigated apoptotic effects of XT5. Treatment of K562 cell lines with a combination of XT5 and
imatinib-XT5 increased cytotoxicity, the Annexin V binding and caspase 3/7 activation. In addition to apoptosis
assays, we observed an increase in the expression levels of the pro-apoptotic (BAX, BAD and BIM) genes in
XT5 treated K562R and K562S cells. Molecular modelling experiments showed that XT5 showed hydrogenbonding
interactions with important amino acids of BCR-ABL kinase receptor; however XT2B did not show
any hydrogen bond interaction.
Conclusion: Our results indicate that XT5 could be a potential candidate to be used as a new anticancer drug
and XT5 combination with imatinib as an alternate treatment strategy for overcoming imatinib resistance.
