Betulinic Acid–Doxorubicin-Drug combination induced apoptotic death via ROS stimulation in a relapsed AML MOLM-13 cell model

Vu, Milan ORCID logoORCID:, Kassouf, Nick ORCID logoORCID: and Appiah, Sandra S. ORCID logoORCID: (2021) Betulinic Acid–Doxorubicin-Drug combination induced apoptotic death via ROS stimulation in a relapsed AML MOLM-13 cell model. Antioxidants, 10 (9) , 1456. ISSN 2076-3921 [Article] (doi:10.3390/antiox10091456)

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In this study, cell death regulation and induction in AML cell line from a relapsed MLL-rearranged cell model (MOLM-13) was investigated with doxorubin (Dox) and betulinic acid (BetA), singly and in combination. CyQUANT Direct® and Annexin V/propidium iodide double staining were used to measure the cytotoxic and cell death induction effects of the compounds, respectively. Reactive oxygen species (ROS) generation was measured using 2′,7′-dichlorofluorescin diacetate staining. Expressions of proteins and genes were examined by Western blot and reverse transcription polymerase chain reaction analysis, respectively. BetA (20 μM) and Dox (1 μM) indicated a synergistic growth inhibitory effect on MOLM-13 cells. The combined drug caused more cells to reside in irreversible late apoptotic stage compared to the single treatments (p < 0.05). Elevation in ROS may be the synergistic mechanism involved in MOLM-13 cell death since ROS can directly disrupt mitochondrial activity. In contrast, in leukaemic U-937 cells, the combination treatments attenuated Dox-induced cell death. Dox and the drug combination selectively reduced (p < 0.05) a recently reported anti-apoptotic Bcl-2 protein isoform p15-20-Bcl-2 in MOLM-13 by our group, without affecting the usually reported p26-Bcl-2-α. Further studies using known inhibitors of apoptosis are required to confirm the potential of Dox-BetA combination to modulate these pathways.

Item Type: Article
Additional Information: This article belongs to the Special Issue Reactive Oxygen Species (ROS), Haematopoiesis and Leukaemia
Keywords (uncontrolled): acute myeloid leukaemia; doxorubicin; betulinic acid; apoptosis; drug combination; B-cell lymphoma 2 family of proteins; reactive oxygen species
Research Areas: A. > Business School > Leadership, Work and Organisations > Diversity and Gender group
A. > School of Science and Technology > Natural Sciences > Biomarkers for Cancer group
Item ID: 33836
Notes on copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (
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Depositing User: Sandra Appiah
Date Deposited: 14 Sep 2021 12:23
Last Modified: 12 Oct 2021 09:58

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