三阴性乳腺癌人类白细胞抗原-G抑制T细胞免疫功能机制的研究  

Mechanism of human leukocyte antigen-G inhibiting T cell immune function in triple-negative breast cancer

作　　者：李晓诗 罗琴 周庆[1] 钟科[1] 蒋安科[1] 夏林玉 胡清林[1] LI Xiaoshi;LUO Qin;ZHOU Qing;ZHONG Ke;JIANG Anke;XIA Linyu;HU Qinglin(Department of Thyroid and Breast Surgery,The First Affiliated Hospital of Chengdu Medical College,Chengdu,Sichuan 610500,China)

机构地区：[1]成都医学院第一附属医院甲乳外科,四川成都610500

出　　处：《中国临床研究》2024年第10期1499-1505,共7页Chinese Journal of Clinical Research

基　　金：成都医学院校基金项目(CYZ19-20)。

摘　　要：目的研究三阴性乳腺癌(TNBC)细胞中人类白细胞抗原-G(HLA-G)表达与T细胞活化发挥免疫抑制功能的机制。方法在三组乳腺癌细胞系(MDA-MB-231,HCC1937,MDA-MB-468)中,实时定量聚合酶链式反应(RT-qPCR)、Western blot检测并筛选出HLA-G高表达,纤维蛋白原样蛋白1(FGL1)、程序性死亡因子配体1(PD-L1)、唾液酸结合免疫球蛋白样凝集素15(SIGLEC15)低表达的乳腺癌细胞系为HCC1937;将HCC1937细胞分为NC组(对照组)与KD组(敲减组),进行RNA干扰(RNAi)慢病毒载体构建并转染KD组细胞以敲减HLA-G基因,且进行验证。HCC1937细胞分别进行HLA-G抗体封闭/未封闭处理,Jurkat细胞分别进行CD3、CD28激活/未激活处理,两种细胞共培养分为8组:NC^(--)组、KD^(--)组、NC^(+-)组、KD^(+-)组、NC^(-+)组、KD^(-+)组、NC^(++)组和KD^(++)组;细胞划痕实验、MTT法、ELISA法分别检测各组细胞迁移率、增殖率以及细胞上清液的干扰素(IFN)-γ、白细胞介素(IL)-2水平。结果RT-qPCR提示HCC1937细胞HLA-G基因敲减成功;KD^(++)组迁移率较NC^(++)组显著增高(P<0.05),但每对共培养组间细胞增殖率差异无统计学意义(P>0.05);KD^(++)组的IFN-γ浓度较NC^(++)组、NC^(-+)组显著降低(P<0.05),但每对共培养组间IL-2浓度差异无统计学意义(P>0.05)。结论TNBC细胞HLA-G可能促进活性T细胞分泌IFN-γ,HLA-G降低后IFN-γ抑制T细胞免疫功能,促进肿瘤转移。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 HCC1937,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)-γand 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-γin 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-γby activing T cells,and IFN-γcan inhibit the immune function of T cells and promote tumor metastasis afte

关 键 词：三阴性乳腺癌 人类白细胞抗原-G T细胞 干扰素-Γ 白细胞介素-2 免疫功能 

分 类 号：R737.9[医药卫生—肿瘤]