Study On The Mechanism Of HDACi Combined With PI3Ki In Acute T Lymphoblastic Leukemia Cell Line Jurkat

Objective: To investigate the effect,synergism and mechanism of histone deacetylase inhibitor(HDACi)chidamide(Chi)and phosphatidylinositol 3kinase inhibitor(PI3Ki)linperlisib(YY-20394)on acute T cell lymphoblastic leukemia(T-ALL)cells.Methods:The Jurkat cell line was treated with different concentrations of chidamide and YY-20394 single drug or combination of two drugs,and the proliferative capacity of the cells after 24,48 and 72 hours of treatment was measured by CCK8 method,and the synergistic effect of the combination group was analyzed.After 48 hours of treatment,the apoptosis rate and cycle distribution of the blank control group,single drug group and combination group were detected by flow cytometry,and the apoptosis of each group was further verified by Tunel fluorescence staining.Finally,use Western-Blotting to detect the expression level of PI3 K,AKT,p-AKT,m-TOR,p-m-TOR,β-actin.Results:The inhibition rate of Jurkat cell proliferation was gradually increased with the increase of concentration and the prolong of time,which was detected by CCK8,with a dose-dependent and time-dependent manner.The combination of the two drugs has synergistic effect on Jurkat cells within a certain concentration range.After 48 hours of treatment,the apoptosis rate and cycle arrest phenomenon of the G0/G1 phase corresponding to the two single drugs and the combination drugs increased with the increase of concentration by flow cytometry.Tunel fluorescence staining also confirmed that the apoptosis of Jurkat cells was more obvious in combination group than the two single drug group.Western blotting assay showed that the expression of AKT and mTOR in the chidamide group was lower than that in the untreated group,and the p-AKT(S473/T308)decreased significantly after treatment.PI3 K,AKT,p-AKT(S473/T308),mTOR,and p-mTOR were significantly down-regulated after single linperlisib treatment and combination group,among which the effect of phosphorylation of the AKT protein was the most obvious.It is worth mentioning that the combination of the two drugs has a more obvious effect on down-regulation of mTOR.Conclusion:Chidamide and YY-20394 either can inhibit acute T lymphoblastic leukemia cells.The combination of the two drugs significantly inhibits the proliferation of T-ALL cell line Jurkat than the single drug,and more significantly promotes apoptosis and cell cycle arrest than the single drug.Chidamide and YY-20394 inhibit the proliferation of T-ALL cell line Jurkat,promote its apoptosis and cell cycle arrest by down-regulating the protein expression of PI3K/AKT/mTOR signal pathway,especially p-AKT,mTOR and p-mTOR.