Lyciumbarbarum polysaccharides inhibit gallbladder cancer cell proliferation,invasion,and immune escape by downregulating the expression of PD-L1
Objective Objective To investigate the effects of lyciumbarbarum polysaccharide(LBP)on the proliferation,metastasis and immune escape of gallbladder carcinoma cells.Methods Gallbladder cancer cells QBC939 and SGC-996 were divided into LBP group,Control group,LBP+pcDNA group,and LBP+PD-L1 group;PBMCs co-incubated with PHA were denoted as PHA group,and PBMCs not co-incubated were denoted as NC group.In addition,QBC939 and SGC-996 cells in the above group were co-incubated with PBMCs,respectively,and divided into PHA+Control group,PHA+LBP+pcDNA group,and PHA+LBP+PD-L1 group.CCK-8,flow cytometry and Transwell assays were used to detect cell proliferation,apoptosis and invasion.Peripheral blood of healthy donors was collected,and peripheral blood mononuclear cells(PBMCs)were isolated by Ficoll-Conray method.The release of IFN-γ in PBMCs induced by phytohemagglutinin(PHA)was detected by ELISA kits.The expression of programmed death ligand 1(PD-L1)was detected by qRT-PCR and western blot.Results Compared with Control group,QBC939 and SGC-996 cells in LBP group showed decreased proliferation(48.00±8.00 vs.100±9.17;39.67±4.51 vs.100±7.00)and invasion(61.33±8.63 vs.110.33±12.50;51.00±8.00 vs.118.67±14.01),and increased apoptosis[(21.10±2.03)%vs.(6.67±1.31)%;(24.30±2.11)%vs.(6.20±1.60)%](P＜0.05).In addition,compared with the PHA group,the IFN-γ content in the PHA+Control group was decreased(2.37±0.15 vs.0.76±0.12,3.11±0.25 vs.1.04±0.19;P＜0.05);Compared with the PHA+Control group,the IFN-γ content in PHA+LBP group was increased(1.84±0.12 vs.0.76±0.12,2.52±0.20 vs.1.04±0.19;P＜0.05).Compared with the Control group,PD-L1 mRNA(1.00±0.07 vs.0.30±0.08,1.00±0.09 vs.0.21±0.07)and protein(1.00±0.11 vs.0.47±0.08,1.00±0.08 vs.0.32±0.07)levels in QBC939 or SGC-996 cells of the LBP group were decreased(P＜0.05).Compared with LBP+pcDNA group,QBC939 or SGC-996 cells in LBP+PD-L1 group had increased cell viability[(48.33±7.02)%vs.(86.00±6.56)%;(40.33±7.02)%vs.(79.33±7.03)%],decreased apoptosis rate[(20.36±3.01)%vs.(13.30±2.01)%;(24.50±1.93)%vs.(14.30±1.77)%]and enhanced cell invasion(56.67±10.01 vs.90.33±9.61;48.33±9.61 vs.95.67±9.50;P＜0.05);Relative to PHA+LPB+pcDNA group,PHA+LPB+PD-L1 group showed decreased IFN-γ levels(1.86±0.16 vs.0.77±0.12,P＜0.05).Conclusion LBP could inhibit the proliferation,invasion and immune escape of gallbladder cancer cells by down-regulating the expression of PD-L1.
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