Effects Of Zanubrutinib Combined With Imatinib On Proliferatio And Apoptosis Of SUP-B15 Cells And Its Related Mechanism

Objective: In this study,zanubrutinib(BGB-3111)and imatinib(IM)were used alone and in combination in Philadelphia chromosome positive acute B-lymphoblastic leukemia cell line SUP-B15,to explore its effects on proliferation and apoptosis and possible mechanism of action,so as to find a new treatment for Ph+ B-ALL and provide a theoretical basis for its clinical application.Methods: 1.The BGB-3111,IM monohydrate and the two drugs were assayed using MTT method to SUP-B15 cells 24 h,48 h,72 h,and the proliferation inhibition was observed.2.Flow cytometry detection BGB-3111,IM monoesion and two phases were combined to SUP-B15 cell strains 48 h,and their apoptosis was observed.3.At 48 hours of time,BGB-3111,IM monohydrate and two drugs were combined with SUP-B15 cell strains,and BCR-ABL,BTK,AKTMRNA were expressed by q-PCR detection.4.At 48 hours of time point,BGB-3111,IM monohydrate and two drugs were combined to SUP-B15 cell strains,and Western blot was used to detect BCR-ABL,BTK,P-Akt,Akt and other protein expression levels.Results:1.MTT method results: As BGB-3111,IM acts in the time of cells,the concentration increases,the increase in the proliferation inhibition rate of cells,and the dose-time dependence.The difference was statistically significant.Through the Jinkic formula,2.5μmol/L BGB-3111+10μmol/L IM 、 10μmol/L BGB-3111+5μmol/L IM 、10μmol/L BGB-3111+10μmol/L IM in the 24 h and 48 h synergies 1.15,where 10μmol/L BGB-3111+5μmol/L IM at 48 h,synergistic is 1.23> 1.15,and the strongest effect is the strongest.Results: BGB-3111 and IM have a synergistic proliferation inhibitory effect on SUP-B15 cells.2.FCM results showed that the average apoptosis rate(1.51 ± 0.87)% in the control group,the average apoptosis rate(8.05 ± 2.02)%,IM(5μmol/L)single The average apoptotic rate(12.07 ± 2.86)% of the pharmaceutical group(10 μmol/L BGB-3111 combined with 5 μmol/L IM)had an average apoptotic rate(26.11 ± 2.78)%.It can be seen that the apoptotic rate combined with the two phases is greater than the single medicine group and the control group(P <0.05).It is concluded that BGB-3111 and IMinduced apoptosis rates are higher than that of each single medicine group,and is mainly apoptosis.3.QPCR results show: BGB-3111(10 μmol/L)single medicine group and IM(5 μmol/L)monovalent drug group and two pharmaceutical joints for SUP-B15 cell strain 48 h,BCR-ABL,AKT,BTKm RNA The expression level is significantly lower than that of the control group,and the expression of AKTm RNA in the combined group of the two pharmaceutical group is significantly lower than that of each single pharmaceutical group.4.Western blot shows: BGB-3111(10μmol/L)single medicine group and IM(5 μmol/L)monhater group and two pharmaceutical joint groups act on SUP-B15 cell strain 48 h,The expression level of BCR-ABL,P-AKT,AKT and BTK protein was lower than that of the control group,and the expression of the P-AKT,AKT protein in the combined group was lower than that of each single pharmaceutical group,and the difference was statistically significant(P <0.05).Conclusion: BGB-3111,IM monohydrate and two drugs are combined to SUP-B15 cell lines,which have proliferative inhibition,induced apoptosis,and consider synergistic effects,possibly with BGB-3111 and IM to inhibit the key on BCR signaling paths.Kinase BTK,BCR-ABL,etc.,further synergies to inhibit the downstream PI3K/AKT signaling pathway.