Bioinformatics-Guided Discovery of miRNAs Involved in Apoptosis Modulated by Parthenolide Combined with Vincristine in The NALM6 Cell Line

Document Type : Original Article

Authors

1 Division of Hematology and Blood bank, Department of Medical Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

2 Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

3 Division of Biochemistry, Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, Iran

4 Department of Biochemistry and Molecular Medicine, University of Montreal, Montreal, Canada

Abstract

Objective: Acute lymphoblastic leukemia (ALL) is a highly heterogeneous leukemia. Despite the current improvement in
conventional chemotherapy and high survival rates, the outcomes remain challenging. Sesquiterpen extracted from the
Tanacetum parthenium, parthenolide, is a potential anticancer agent that can modulate the expression of miRNAs and induce
apoptosis. The objective of this study was to investigate the effect of parthenolide in combination with vincristine and alone on
the apoptosis rate and expression of miR-125b-5p, miR-181b-5p, and miR-17-5p in the NALM6 cell line.
Materials and Methods: In this experimental study, cell viability and metabolic activity were determined through MTT
assay and PI staining. Flow cytometry was applied to evaluate the rate of apoptosis. The expression of miRNAs was
assessed using real-time polymerase chain reaction. Bioinformatic analyses, including Cytoscape, RNAhybrid, and
signaling pathway analysis were employed to investigate the association of miR-17-5p, miR-181b-5p and miR-125b-
5p with apoptosis. Further, molecular docking served to validate the modulation of these miRNAs by parthenolide and
vincristine treatment.
Results: The MTT assay indicated that 7.7 μM of parthenolide decreased the metabolic activity to 50% after 48 hours. PI
staining analysis indicated that at concentrations below the half maximal inhibitory concentration, parthenolide caused
50% cell death. Flow cytometric analysis indicated that parthenolide (1.925 μM) in combination with vincristine (1.2 nM)
induced apoptosis in 83.2% of the cells. Real-time quantitative reverse transcription polymerase chain reaction (qRTPCR)
analysis showed significant changes in the expression levels of miR-17-5p, miR-125b-5p, and miR-181b-5p.
Moreover, the combination therapy downregulated the expression of miRNAs significantly. This was consistent with our
bioinformatic analysis demonstrating that the studied miRNAs are regulators of apoptosis. Finally, molecular docking
validated the modulation of the miRNAs by parthenolide and vincristine.
Conclusion: Parthenolide in combination with vincristine triggers apoptosis at a high rate in the NALM6 cell line.
Moreover, this combination therapy can decrease the expression of miR-17-5p, miR-181b-5p, and miR-125b-5p.

Keywords

Main Subjects

References

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Cell Journal (Yakhteh)
Volume 26, Issue 2 - Serial Number 121
February 2024
Pages 139-149

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