Pinostrobin induces acute leukemia cell apoptosis via the regulation of miR-410-5p and SFRP5

Aims: This study attempted to explore the mechanisms involved in pinostrobin (PN)-mediated acute leukemia cell Apoptosis regulated by miR-410-5p.

Material and methods: NB4 and MOLT-4 cells were cultured and treated with PN at the IC50 concentration. Apoptosis was examined by Annexin V-FITC/PI staining. RT-qPCR was used to measure the expression of Caspase-3, Bak, Bcl-W, and Mcl-1. The target protein of PN was identified using LC-MS/MS followed by bioinformatic analysis. TargetScan, DIANA, and miRDB were used for the prediction of miRNAs involved in the PN-induced Apoptosis mechanism. miRNA mimic transfection, RT-qPCR, and western blot analysis were performed to evaluate the regulatory effect of miRNA on its target and the involvement of miRNA in Apoptosis induction by PN. In addition, the synergistic effect of PN and daunorubicin (DNR) were investigated by using the MTT assay.

Key findings: The results showed that PN reduced cell viability and induced Apoptosis in both leukemia cell lines. From the LC-MS/MS and bioinformatics analysis, SFRP5 and miR-410-5p were selected as a potential PN target protein and miRNA, respectively. After miRNA mimic transfection, miR-410-5p, which is an onco-miRNA, was decreased and led to increased Apoptosis in both cell lines, indicating that this miRNA is involved in PN-mediated Apoptosis mechanisms. Moreover, PN demonstrated a synergistic effect with DNR, suggesting that PN may be used in combination with conventional chemotherapy drugs.

Significance: PN regulates the expression of miR-410-5p and SFRP5 to promote Apoptosis in acute leukemia cells. It could be developed as an alternative treatment for leukemia in the future.

Leukemia; Pinostrobin; SFRP5; miRNAs.