Effects of cyclin-dependent kinase 4/6 inhibitors on B-cell maturation antigen and programmed death-ligand 1 in myeloma cells
Objective To investigate the influences of cyclin-dependent kinase 4/6(CDK4/6)inhibitors on the expressions of multiple myeloma(MM)cellular immunotherapy-associated molecules B-cell maturation antigen(BCMA)and programmed death-ligand 1(PD-L1)and on the apoptosis of MM cells and cells in normal microenvironment.Methods H929 cells in logarithmic growth phase were divided into abemaciclib,palbociclib and DMSO groups,and treated with 2 μmol/L abemaciclib,5 μmol/L palbociclib and equivalent volume of DMSO in cell culture medium,respectively.The relative mean fluorescence intensities of BCMA and PD-L1 were detected by flow cytometry on day 1,3 and 5 of culture in three groups.The bone marrow was collected from 3 MM patients diagnosed and treated in Blood Diseases Hospital of Chinese Academy of Medical Sciences from January,2020 to June,2023,and the bone marrow mononuclear cells were separated by using lymphocyte isolation solution,and divided into abemaciclib,palbociclib and DMSO groups,which were treated with 1 μmol/L abemaciclib,10 μmol/L palbociclib,and equivalent volume of DMSO,respectively.Flow cytometry was used to detect the relative cell survival rates of CD138+tumor cells and CD138-cells in normal microenvironment.Results The relative mean fluorescence intensities of BCMA were higher in palbociclib group(1.37±0.04,2.36±0.11,3.99±0.17)than those in abemaciclib group(0.69±0.02,1.03±0.05,1.69±0.06)on day 1,3 and 5 of culture(P＜0.05),and were higher in palbociclib group than those in DMSO group(1.00±0.02,1.00±0.05)on day 3 and 5(P＜0.05),while there was no significant difference between palbociclib group and DMSO group on day 1(P＞0.05).In abemaciclib group,the relative mean fluorescence intensity of BCMA was higher than that in DMSO group on day 5(P＜0.05),and showed no significant difference on day 1 and 3 compared with DMSO group(P＞0.05).The relative mean fluorescence intensities of BCMA increased sequentially in abemaciclib group and palbociclib group on day 1,3 and 5(P＜0.05).On day 1,3 and 5,the relative mean fluorescence intensities of PD-L1 were higher in abemaciclib group(1.30±0.02,3.21±0.10,2.28±0.17)and palbociclib group(1.28±0.12,2.81±0.38,2.00±0.16)than those in DMSO group(1.00±0.09,1.00±0.06,1.00±0.14)(P＜0.05),and showed no significant differences between abemaciclib group and palbociclib group(P＞0.05).The relative mean fluorescence intensities of PD-L1 were higher on day 3 in both abemaciclib and palbociclib groups than those on day 1 and 5(P＜0.05),and were higher on day 5 than those on day 1(P＜0.05).After 24 and 48 h of treatment,the relative cell survival rates of CD138+were lower in abemaciclib group and palbociclib group than those in DMSO group(P＜0.05),and lower in palbociclib group than those in abemaciclib group(P＜0.05).The relative cell survival rate of CD138-was lower in palbociclib group than those in DMSO group and abemaciclib group after treatment for 48 h(P＜0.05),and showed no significant difference in palbociclib group after treatment for 24 h compared with DMSO group and abemaciclib group(P＞0.05).Conclusion CDK4/6 inhibitors up-regulate BCMA and PD-L1 expressions in MM cells and selectively kill MM cells,with no significant influence on the cells in normal microenvironment.
NETL
NSTL
万方数据
