Phyllanthin inhibits MOLT‐4 leukemic cancer cell growth and induces apoptosis through the inhibition of AKT and JNK signaling pathway

Abstract Among cancers, leukemia is a multistep progression that involves genetic modifications of normal hematopoietic progenitor cells to cancerous cells. In recent times, leukemia cases and their mortality rate have increased rapidly. Therefore, the immense need for a therapeutic approach is crucial that can control this type of cancer. Phyllanthin is a lignan compound constituent from the Phyllanthus species and has numerous beneficial effects as a dietary component. The present study aims to determine the impact of phyllanthin on the MOLT‐4 cytotoxic effect. MOLT‐4 cells and MS‐5 cells were cultured at different concentrations of phyllanthin (5, 10, 25, 50, 75, and 100 μM/ml), and the viability was assessed by the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide method. The level of reactive oxygen species, the membrane potential of mitochondria, apoptosis by 2',7'‐dichlorofluorescin‐diacetate (DCF‐DA), rhodamine, acridine orange (AO)/ethidium bromide (EB), 4′,6‐diamidino‐2‐phenylindole (DAPI)/propidium iodide (PI) staining, gene expression of signaling molecules, and protein levels were assessed by reverse‐transcription polymerase chain reaction and western blot analysis. Phyllanthin did not show toxicity toward MS‐5 cells and significantly decreased the cell viability of MOLT‐4 cells with an IC 50 value of 25 µM/ml. Also, phyllanthin induced the production of reactive oxygen species and led to the loss of mitochondrial membrane potential. AO/EB and DAPI/PI staining fluorescent image confirmed the induction of apoptosis by phyllanthin treatment. The messenger RNA (mRNA) expression of cell cycle regulator cyclin D1, antiapoptotic gene Bcl‐2, NF‐κB, and TNF‐α decreased, but the proapoptotic Bax mRNA expression was increased. The phosphorylated protein levels of p‐PI3K1/2, p‐ERK1/2, and p‐AKT were decreased, whereas the levels of p‐p38 and p‐JNKT1/2 increased. Our results confirmed that phyllanthin inhibits the MOLT‐4 cells, increases apoptosis, and inhibits MOLT‐4 migration and cell invasion. Therefore, phyllanthin can be used as a potential target for leukemia treatment.