The effect of Ganoderic Acid A on miR-17-5p and miR-181b expression level and apoptosis induction in human leukemia Nalm-6 cells

Faezeh  Mortazavie; Simin  Taheri; Parisa  Tandel; Farahnaz  Zare; Gholmhossein  Tamaddon

doi:10.18502/ijpho.v12i3.10058

Faezeh Mortazavie Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Simin Taheri Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Parisa Tandel Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Farahnaz Zare Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Gholmhossein Tamaddon Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.

DOI: https://doi.org/10.18502/ijpho.v12i3.10058

Keywords: Apoptosis, Ganoderic Acid A, MiR-17-5p, MiR-181b, Nalm-6 cells

Abstract

Background: In various cancers, Ganoderic Acid A (GAA), an active triterpenoid derived from Ganoderma lucidum, has been proved to show potent anti-tumor effects. However, the possible impacts of GAA on the human leukemia cell line (Nalm-6) are not fully elucidated. Therefore, this research aimed to study the antineoplastic effect of GAA on Nalm-6 cells.

Materials and Methods: In this laboratory trial study, Nalm6 cells were cultured in vitro and treated with different doses of GAA (25, 50, 100, 200, and 400 μg/mL) for 24, 48, and 72 hours. The optimal treatment concentration of GAA was determined by the MTT assay. Flow cytometry was used to determine the death of Nalm-6 cells caused by GAA treatment by utilizing FITC-conjugated propidium iodide (PI) and annexin V staining. After incubation, the expression levels of miR-17-5p and miR-181b were monitored using real-time polymerase chain reaction (PCR).

Results: Based on the half-maximal inhibitory concentration (IC50) measurements of the MTT assay, the optimal treatment concentration of GAA was 140 μg/mL (in a dose and time-dependent manner, p<0.0001). The GAA treatment was selectively toxic to the leukemia Nalm-6 cells and could remarkably induce cell apoptosis (p<0.0001). Besides, GAA downregulated the expression of miR-17-5p and miR-181b in the Nalm-6 cells compared with the untreated cells (P=0.0067 and P=0.0014, respectively).

Conclusions: Based on the present findings, GAA merits further investigation as a promising natural reagent for treating hematologic malignancies.