The pleiotropic effects of fisetin and hesperetin on human acute promyelocytic leukemia cells are mediated through apoptosis, cell cycle arrest, and alterations in signaling networks

Abstract

Fisetin and hesperetin, flavonoids from various plants, have several pharmaceutical activities including antioxidative, anti-inflammatory, and anticancer effects. However, studies elucidating the role and the mechanism(s) of action of fisetin and hesperetin in acute promyelocytic leukemia are absent. In this study, we investigated the mechanism of the antiproliferative and apoptotic actions exerted by fisetin and hesperetin on human HL60 acute promyelocytic leukemia cells. The viability of HL60 cells was evaluated using the MTT assay, apoptosis by annexin V/propidium iodide (PI) staining and cell cycle distribution using flow cytometry, and changes in caspase-3 enzyme activity and mitochondrial transmembrane potential. Moreover, we performed wholegenome microarray gene expression analysis to reveal genes affected by fisetin and hesperetin that can be important for developing of future targeted therapy. Based on data obtained from microarray analysis, we also described biological networks modulated after fisetin and hesperetin treatment by KEGG and IPA analysis. Fisetin and hesperetin treatment showed a concentration-and time-dependent inhibition of proliferation and induced G2/M arrest for both agents and G0/G1 arrest for hesperetin at only the highest concentrations. There was a disruption of mitochondrial membrane potential together with increased caspase-3 activity. Furthermore, fisetin-and hesperetin-triggered apoptosis was confirmed by annexin V/PI analysis. The microarray gene profiling analysis revealed some important biological pathways including mitogen-activated protein kinases (MAPK) and inhibitor of DNA binding (ID) signaling pathways altered by fisetin and hesperetin treatment as well as gave a list of genes modulated ≥2-fold involved in cell proliferation, cell division, and apoptosis. Altogether, data suggested that fisetin and hesperetin have anticancer properties and deserve further investigation.

Dietary flavonoid fisetin for cancer prevention and treatment

“…Fisetin and hesperetin induced apoptosis along with inhibited cell proliferation, induced G2/M arrest, disrupted the mitochondrial potential, and increased caspase‐3 activity. Microarray gene profiling of treated cells revealed some important biological pathways including MAPK, DNA binding signaling pathways and genes involved in cell proliferation, division, and apoptosis . Another similar study demonstrated the effect of fisetin and hesperetin combination on human K562 CML cells.…”

“…Combination treatment significantly modulated genes involved in cell proliferation, cell division, apoptosis, cell cycle, and various other cellular processes such as replication, transcription, and translation. Microarray gene profiling analysis revealed genes involved in JAK/STAT pathway, KIT receptor, and growth hormone receptor signaling to be potential candidates of fisetin‐hesperetin combination for targeted CML therapy .…”

Lead Phytochemicals for Anticancer Drug Development

“…mitogen activated protein kinases (MAPK), NF-κB, JAK/STAT, PI3K/Akt, Wnt, and mTOR pathways (Adan and Baran, 2015). Genistein, the isoflavone of soybean exerts its anticancer effects via inhibition of NF-κB and Akt signaling pathways (Li et al, 2012).…”

Leukemia therapy by flavonoids: Future and involved mechanisms

“…Likewise, in flavonoid‐treated Kasumi‐1 cells, leukemic cell line with an 8; 21 chromosome translocation, activated form of caspase 3, 9, and PARP improved meaningfully, and consequently apoptosis rate of target cells augmented noticeably (B. Wang et al, ). As well, it was shown that flavonoid, hespertin, and fisetin, increased activated form of the caspase 3 in flavonoid‐treated acute promyelocytic leukemia (APL) cells (Adan & Baran, ). In addition, flavonoids in AML cells carrying nucleophosmin 1 (NPM1) mutation‐induced apoptosis via the activation of caspase 8.…”