STAT3磷酸化调控乳腺癌髓系来源抑制细胞介导T细胞免疫抑制的机制研究  

Activation of STAT3 promoting immunosuppressive effect on T cells in MDSCs of breast cancer

作　　者：王越[1] 任秀宝[1] 李慧[1] 曹水[1] 任宝柱[1] 于文文[1] 张澎[1] 齐静[1] 于津浦[1] 

机构地区：[1]天津医科大学肿瘤医院免疫室,国家肿瘤临床医学研究中心,天津市肿瘤免疫与生物治疗重点实验室,天津市300060

出　　处：《中国肿瘤临床》2013年第17期1016-1019,共4页Chinese Journal of Clinical Oncology

基　　金：国家重点基础研究发展计划(编号:2012CB9333004);国家自然科学基金项目(编号:81072159);天津市高等学校科技发展基金计划项目(编号:20090133)资助~~

摘　　要：目的:检测STAT3在乳腺癌MDSCs中的磷酸化状态及其对MDSCs免疫抑制活性的影响。方法:收集脐血单个核细胞,免疫磁珠的方法分选其中CD33+细胞,体外与乳腺癌细胞系MDA-MB-231共孵育诱导MDSC生成。Western blot方法分别检测IDO和STATs的表达与磷酸化情况。MDSCs与健康供者外周血T淋巴细胞共孵育,分别加入1-MT和JSI-124来抑制IDO功能和STAT3磷酸化,利用MTT实验和ELISA检测各组T细胞的增殖和细胞因子分泌。结果:Western blot检测发现体外诱导的MDSCs中IDO表达明显增加,同时伴STAT3磷酸化水平升高。加入JSI-124后pSTAT3和IDO表达明显降低。MTT实验中MDSCs明显抑制T细胞增殖,加用IDO特异性抑制剂1-MT或STAT3抑制剂JSI-124后T细胞增殖抑制明显改善(P<0.05),且1-MT组和JSI-124组之间差异无统计学意义。ELISA结果显示MDSCs显著抑制T细胞分泌IFN-γ,促进TGF-β、IL-10释放(P<0.05)。而加用1-MT或JSI-124后,IFN-γ分泌水平升高,而TGF-β、IL-10分泌水平降低(P<0.05),而1-MT组和JSI-124组之间差异无统计学意义。结论:MDSCs中磷酸化STAT3水平升高导致IDO过表达;STAT3的特异性抑制剂JSI-124可以逆转MDSCs对T细胞增殖和Th1类因子分泌的抑制作用。Objective：To explore the status of STAT3 phosphorylation in myeloid-derived suppressor cells （MDSCs） of breast cancer and its function in the immunosuppressive effect of MDSCs on proliferation and cytokine secretion of T cells. Methods：CCD33＋cells were isolated from healthy umbilical cord, blood-derived, peripheral blood mononuclear cells and were co-cultured with breast cancer cell line MDA-MB-231 in vitro using Transwell plates to induce MDSCs. The untreated CD33＋cells were used as con-trols. Idoxuridine （IDO） suppressor expression and STAT3 phosphorylation were examined using Western blot assay. The proliferation and cytokine secretion of T cells, which were co-cultured with MDSCs, were determined by methyl thiazol tetrazolium assay and en-zyme-linked immunosorbent assay. 1-MT and JSI-124 were used to investigate the function of IDO and pSTAT3 in MDSC-mediated T cell immunosuppression. Results：The protein levels of IDO and pSTAT3 in MDSCs were significantly upregulated. MDSCs obviously suppressed T-cell proliferation, which was reversed by 1-MT or JSI-124 （P〈0.05）. MDSCs could promote TGF-βand IL-10 secretions, but could also remarkably inhibit IFN-γsecretion （P〈0.05）. After incubation with 1-MT or JSI-124, the increase in TGF-βand IL-10, as well as the decrease in IFN-γ, was significantly reversed. Conclusion：The upregulated pSTAT3 induced the IDO increase in MDSCs. JSI-124 can block MDSC-mediated immunosuppressive effect on T cells in breast cancer.

关 键 词：髓系来源抑制细胞 STAT3 T细胞 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]