Unveiling Novel Insights in Chronic Myeloid Leukemia Cell Lines K562 through in vitro and in silico Approach Targeting of Apoptosis and Oxidative Stress Pathways

Abstract

Introduction: Troxerutin (TR), bioflavanoid derived from rutin posess several biological activity like antioxidant, antinflammatory, antidiabetic and antitumor activity and found to be alternative candidate for cancer therapy. Chronic Myelogenous Leukaemia (CML) is a hematoproliferative disorder caused by uncontrolled myeloid cell division in bone marrow charecterized by Bcr and Abl gene fusion. This study aimed to evaluate the anticancer activity of TR on K562 chronic myeloid leukemia cell line. Materials and Methods: The cytoxocity was assessed by MTT and phase contrast microscopy. DAPI staining was carried out to confirm apoptosis. The antioxidant status was determined by Glutathione (GSH) levels. Molecular docking was carried out to understand the interaction between p53 and TR. Caspase-9 level was quantified by ELISA whereas gene expression of BAX, p53 and Bcl-2 level by qRT-PCR. Results: The cell growth of K562 was reduced in dose dependent manner with rounding and cell shrinkage. Oxidative stress mediated apoptosis was observed by elevated GSH level and nuclear condensation by DAPI. The increased gene expression level of Bax and p53 with decreased level of Bcl-2 was confirmed. Molecular docking analysis depicted binding affinity of TR with p53 confirming its potential role in apoptosis induction. Conclusion: The anticancer activity of troxerutin against K562 cells was induced apoptosis by ROS-mediated oxidative stress and p53-dependent apoptotic pathway supporting therapeutic potential to treat CML.