GSK-J4 induces cell cycle arrest and apoptosis via ER stress and the synergism between GSK-J4 and decitabine in acute myeloid leukemia KG-1a cells

Background: GSK-J4 is the inhibitor of H3K27me3 demethylase. Recent studies demonstrated that GSK-J4 could affect the proliferation and Apoptosis of a variety of Cancer cells. However, the effects and underlying mechanisms of GSK-J4 on the proliferation and Apoptosis of human acute myeloid leukemia (AML) KG-1a cells have not been explored thoroughly.

Methods: The effect of GSK-J4 on cell proliferation was assessed with CCK8, while cell cycle distribution and Apoptosis were analyzed using flow cytometry. The proteins related to cell cycle, cell Apoptosis, endoplastic reticulum (ER) stress and PKC-α/p-Bcl2 pathway were detected by Western blotting. The expression level of PKC-α mRNA was measured by quantitative Real-Time PCR.ER stress inhibitor 4-phenyl butyric acid (4-PBA) was used to explore the role of ER stress in GSK-J4 induced cell-cycle arrest and cell Apoptosis. The combination effects of Decitabine and GSK-J4 on KG-1a cells proliferation and Apoptosis were also evaluated by CCK8, flow cytometry and immunoblot analysis.

Results: GSK-J4 reduced cell viability and arrested cell cycle progression at the S phase by decreasing the expression of CyclinD1 and CyclinA2 and increasing that of P21. Moreover, GSK-J4 enhanced the expression of apoptosis-related proteins (cle-caspase-9 and Bax) and inhibited PKC-a/p-Bcl2 pathway to promote cell Apoptosis. In addition, ER stress-related proteins (caspase-12, GRP78 and ATF4) were increased markedly after exposure to GSK-J4. The effects of GSK-J4 on cell cycle, Apoptosis and PKC-a/p-Bcl2 pathway were attenuated after treatment with ER stress inhibitor. Furthermore, decitabine could significantly inhibit the proliferation and induce the Apoptosis of KG-1a cells after combined treatment with GSK-J4.

Conclusion: Taken together, this study provided evidence that ER stress could regulate the process of GSK-J4-induced cell cycle arrest, cell Apoptosis and PKC-α/p-bcl2 pathway inhibition and demonstrated a potential combinatory effect of decitabine and GSK-J4 on leukemic cell proliferation and Apoptosis.