Inhibition of Autophagy Augments the Anticancer Activity of KPT-330 in Mantle Cell Lymphoma Cells
Objective:To investigate the influence of novel CRM1 inhibitor KPT-330 on the autophagy of mantle cell lymphoma(MCL)cells,and effect of KPT-330 on the prolifiration of MCL cells in the presence or absence of autophagy inhibitor.Methods:CCK-8 assay was used to detect the effect of KPT-330 on MCL cell lines Z-138,Jeko-1,Granta-519,Rec-1.The effect of KPT-330 on autophagy features were determined by detecting acidic vesicular organelles(AVO)by MDC staining under fluorescence microscope and detecting protein expression of LC3B-Ⅱ assessed by Western blot.Further combined application of lysosomal inhibitor Chloroquine(CQ)was used to observe its effect on the increase of LC3B-Ⅱ caused by KPT-330.CalcuSyn 2.0 software was used to detected the Combination index(CI)of KPT-330 combined with CQ.Results:The proliferation of MCL cell lines(Z-138,Jeko-1,Grant-519,Rec-1)could be inhibited by KPT-330 in a dose-dependent manner(r=0.930,0.946,0.691,0.968 respectively).The number of acidic vesicular organelles(AVO)and the expression of LC3B-Ⅱ were increased in KPT-330 treated Jeko-1 and Granta-519 cells in a dose-dependent manner(rJeko-1=0.993,rGranta-519=0.971).LC3B-Ⅱ protein amounts still increased upon KPT-330 treatment with the existence of lysosomal inhibitor CQ in Jeko-1 and Granta-519 cells,which was higher than CQ or KPT-330 single drug group.The combination of KPT-330 and CQ produced the synergistic effects on cells proliferation inhibition with CalcuSyn 2.0 analysis.Conclusion:KPT-330 can inhibit MCL cell proliferation and induce autophagy.KPT-330 combined with autophagy inhibitor CQ could produce synergistic anti MCL effects,providing experimental basis for clinical combination therapy.
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