Thymoquinone Inhibits JAK/STAT and PI3K/Akt/ mTOR Signaling Pathways in MV4-11 and K562 Myeloid Leukemia Cells

Abstract

Constitutive activation of Janus tyrosine kinase-signal transducer and activator of transcription (JAK/STAT) and Phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathways plays a crucial role in the development of acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). Thymoquinone (TQ), one of the main constituents of Nigella sativa, has shown anti-cancer activities in several cancers. However, the inhibitory effect mechanism of TQ on leukemia has not been fully understood. Therefore, this study aimed to investigate the effect of TQ on JAK/STAT and PI3K/Akt/mTOR pathways in MV4-11 AML cells and K562 CML cells. FLT3-ITD positive MV4-11 cells and BCR-ABL positive K562 cells were treated with TQ. Cytotoxicity assay was assessed using WSTs-8 kit. The expression of the target genes was evaluated using RT-qPCR. The phosphorylation status and the levels of proteins involved in JAK/STAT and PI3K/Akt/mTOR pathways were investigated using Jess western analysis. TQ induced a dose and time dependent inhibition of K562 cells proliferation. TQ significantly downregulated PI3K, Akt, and mTOR and upregulated PTEN expression with a significant inhibition of JAK/STAT and PI3K/Akt/mTOR signaling. In conclusion, TQ reduces the expression of PI3K, Akt, and mTOR genes and enhances the expression of PTEN gene at the mRNA and protein levels. TQ also inhibits JAK/STAT and PI3K/Akt/mTOR pathways, and consequently inhibits proliferation of myeloid leukemia cells, suggesting that TQ has potential anti-leukemic effects on both AML and CML cells.

Thymoquinone Enhances Apoptosis of K562 Chronic Myeloid Leukemia Cells through Hypomethylation of SHP-1 and Inhibition of JAK/STAT Signaling Pathway

“…Additionally, TQ inhibited STAT3 phosphorylation in Multiple myeloma cells [ 47 ] and MV4-11 leukemia cells [ 27 ]. Moreover, in our previously published report, we indicated that TQ had reduced the phosphorylation and the protein levels of JAK2, STAT3, and STAT5 protein intensities and phosphorylation in MV4-11 AML cells [ 48 , 49 ]. The findings of this study are also in agreement with previous findings in which TQ treatment reduced the phosphorylation of JAK2 and STAT3, which consequently led to enhanced apoptosis in SK-MEL-28 melanoma cells [ 50 ].…”

Involvement of Nrf2, JAK and COX pathways in acetaminophen-induced nephropathy: Role of some antioxidants

“…Similarly, the JAK-STAT stimulation in the Helicobacter-pylori-infected gastric epithelial cells was abolished by treatment with α-LA ( Choi et al, 2016 ). Also, TQ was found to induce anticancer effect on myeloid leukemia cells via inhibition JAK/STAT and PI3K/Akt/mTOR pathways ( Al-Rawashde et al, 2022 ).…”

Immune-Stimulating Potential of Lacticaseibacillus rhamnosus LM1019 in RAW 264.7 Cells and Immunosuppressed Mice Induced by Cyclophosphamide

“…The primers used for this experiment are listed in Table 1. The protein expression level changes induced by LM1019 in RAW 264.7 cells were assessed using automated capillary western blot assays with the Jess™ system (ProteinSimple, Bio-Techne, Minneapolis, CA, USA) [34][35][36]. RAW 264.7 cells were seeded at a density of 9 × 10 5 cells/well in a 6-well plate and incubated for 24 h at 37 • C in a 5% CO 2 incubator.…”