Objective To investigate the mechanism of human leukocyte antigen-G(HLA-G)expression and T cells activation in triple-negative breast cancer(TNBC)cells to exert immunosuppressive function.Methods From three groups of breast cancer cell lines(MDA-MB-231,HCC1937,MDA-MB-468),real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot were detected and screened that the breast cancer cell line was HCC 1937,which with high expression of HLA-G,and low expression of fibrinogen-like protein 1(FGL1),programmed death factor ligand 1(PD-L1),as well as sialic acid-binding immunoglobulin-like lectin 15(SIGLEC15).HCC1937 cells were divided into NC group(control group)and KD group(knockdown group),and RNA interference(RNAi)lentiviral vectors were constructed and transfected into KD cells to knock down HLA-G gene,and the verification was carried out.HCC1937 cells were treated with HLA-G antibody blocking/nonblocking,and Jurkat cells were treated with CD3 and CD28 activation/non-activation,respectively,and the two types of cells were co-cultured into 8 groups:NC--,KD--,NC+-,KD+-,NC-+,KD-+,NC++,and KD++groups.Cell scratch assay,MTT and ELISA method were used to detect cell migration rate,proliferation rate,and interferon(IFN)-γ and interleukin(IL)-2 levels in cell supernatants,respectively.Results RT-qPCR showed that the HLA-G gene was successfully knocked down in HCC1937 cells.Compared with the NC++group,the migration rate of the KD++group was significantly increased(P＜0.05),but there was no statistical difference in the cell proliferation rate between each pair of co-culture groups(P＞0.05).The concentration of IFN-γ in the KD++group was lower than that in the NC++group and the NC-+group(P＜0.05),but there was no significant difference in the concentration of IL-2 between each pair of co-culture groups(P＞0.05).Conclusion HLA-G in TNBC cells may promote the secretion of IFN-γ by activing T cells,and IFN-γ can inhibit the immune function of T cells and promote tumor metastasis after HLA-G decreased.
Triple-negative breast cancerHuman leukocyte antigen-GT cellInterferon-γInterleukin-2Immune function
维普
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