Abstract
Myeloid leukemia is one of the major causes of deaths among elderly with very poor prognosis. Due to the adverse effects of existing chemotherapeutic agents, plant-derived components are being screened for their anti-leukemic potential. Momordica charantia (bitter gourd) possesses a variety of therapeutic activities. We have earlier demonstrated anti-leukemic activity of acetone extract of M. charantia seeds. The present study reports purification of differentiation inducing principle(s) from further fractionated seed extract (hexane fraction of the acetone extract, Mc2-Ac-hex) using HL-60 cells. Out of the 5 peak fractions (P1–P5) obtained from normal phase HPLC of the Mc2-Ac-hex, only peak fraction 3 (P3) induced differentiation of HL-60 cells as evident from an increase in NBT-positive cells and increased expression of cell surface marker CD11b. The P3 differentiated the HL-60 cells to granulocytic lineage, established by increased CD15 (granulocytic cell surface marker) expression in the treated cells. Further, possible molecular mechanism and the signalling pathway involved in the differentiation of HL-60 cells were also investigated. Use of specific signalling pathway inhibitors in the differentiation study, and proteome array analysis of the treated cells collectively revealed the involvement the of ERK/MAPK mediated pathway. Partial characterization of the P3 by GC–MS analysis revealed the presence of dibutyl phthalate, and derivatives of 2,5-dihydrofuran to be the highest among the 5 identified compounds. This study thus demonstrated that purified differentiation-inducing principle(s) from M. charantia seed extract induce HL-60 cells to granulocytic lineage through ERK/MAPK signalling pathway.
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All the data used to support the findings of this study are available from the corresponding author upon request.
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Acknowledgements
JS and PP thank the Jawaharlal Nehru University, New Delhi, for the research fellowship. RS and SG acknowledge the University Grants Commission, New Delhi for the research fellowship. AD acknowledges intramural support from the Jawaharlal Nehru University, New Delhi. JS thanks Dr. Ankita Pandey and Dr. Sapna Sharma for their inputs. Authors thank Dr. Swati Tiwari, School of Biotechnology, Jawaharlal Nehru University, New Delhi for fruitful discussions, valuable inputs and access to her lab facility for immunoblotting analysis.
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JS and PP: formal execution, analysis of experiments, data curation, writing—original draft preparation. RS: experimentation and manuscript draft preparation. SG: experimentation and data curation. AD: conceptualization, project administration, and supervision, writing—review, and editing.
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Sharma, J., Prabha, P., Sharma, R. et al. Anti-leukemic principle(s) from Momordica charantia seeds induce differentiation of HL-60 cells through ERK/MAPK signalling pathway. Cytotechnology 74, 591–611 (2022). https://doi.org/10.1007/s10616-022-00547-x
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DOI: https://doi.org/10.1007/s10616-022-00547-x