凝血因子Ⅻ诱导外周血单核细胞分化为卵巢癌腹膜内肿瘤相关巨噬细胞的实验研究  

Coagulation factor Ⅻ induced MO differentiation into a TAM like macrophage subset in ovarian cancer

作　　者：王睿黎[1] 张婷[1] 马政文[2] 王颖[3] 马庆良[1] 徐红[1] 程忠平[4] 汪希鹏[1] 

机构地区：[1]上海交通大学医学院附属仁济医院妇产科,上海200001 [2]上海交通大学医学院基础医学院神经生物教研室 [3]上海交通大学医学院免疫所 [4]上海市杨浦区中心医院妇产科

出　　处：《现代妇产科进展》2010年第4期270-273,277,共5页Progress in Obstetrics and Gynecology

基　　金：国家自然基金资助项目(No:30600672);上海市科委启明星后资助(No:06QH14010)

摘　　要：目的:探讨凝血因子Ⅻ能否诱导外周血单核细胞(MO)分化为卵巢癌(EOC)腹膜微环境中的肿瘤相关巨噬细胞(TAM)。方法:分离正常女性外周血MO,在体外经Ⅻ因子刺激,流式荧光激活细胞分选技术(FACS)检测刺激后CD14、CD68和CD163表达的比例;ELISA检测TNF-α、IL-4、IL-8、TGF-β、IL-10、MMP2等因子的表达;Real-timePCR检测IL-10、IL-8、CCL18、CCR2、TGF-β、CXCR1及CXCR2等细胞因子及相关受体mRNA的表达,添加Ⅻ因子抑制物-C1酯酶抑制剂(C1-INH)后检测相应mRNA转录变化。结果:经Ⅻ因子刺激后,MO表面CD14,CD163表达比例上调[CD14:(57.025±11.135)%,CD163:(1.09±0.21)%],与阴性对照组[CD14:(45.2±5.24)%,CD163:(0.7±0.08)%]相比有统计学差异(P<0.05);经Ⅻ因子刺激后MO分泌IL-8,IL-10及TGF-β分泌表现出时间依赖性,IL-8及TGF-β于刺激后12h分泌达到高峰,IL-10分泌高峰为刺激后3h;刺激后IL-8,IL-10,CXCR2,CCR2以及CCL18mRNA的表达上调,添加C1-INH后,相应mRNA的转录水平下调。结论:凝血因子Ⅻ能诱导外周血MO分化成表型为CD14highCD163highIL-10highCCL18highIL-8high的单核巨噬细胞,类似TAM特征的巨噬细胞亚型,Ⅻ因子可能参与EOC腹膜微环境中浸润的MO分化,从而调控EOC的腹膜转移。Objective：To investigate whether coagulation factor Ⅻ（FⅫ） could induce peripheral blood monocytes（MOs） into a TAM（tumor associated macrophage）-like macrophage in ovarian cancer.Methods：MOs from healthy female donors were culture and stimulated with FⅫ.The following experiments were performed：FACS was used to analyzed the phenotype of MOs（CD14,CD68,CD163）after stimulated with FⅫ.After the isolated MOs were stimulated with FⅫ,the cell culture supernatants were collected,then the expression of cytokines （IL-8、IL-10、TNF-α、IL-4、TGF-βand MMP2） was measured by ELISA.Real-time PCR was used to detect mRNA expression of FⅫ-stimulated MOs,including IL-8、IL-10、CCL18、CCR2、TGF-β、CXCR1 and CXCR2.C1-INH were used as the inhibitor of FⅫ.Results：MOs stimulated by FⅫ maintained higher percentage of CD14[（57.025±11.135）%]and CD163[（1.09±0.21）%] than those of MOs cultured with FⅫ-free medium[CD14：（45.2±5.24）%,CD163：（0.7±0.08）%]（P〈0.05）.In ELISA experiment,IL-8、TGF-β and IL-10 demonstrated a time-dependent expression profile.IL-8 and TGF-β were at maximal production when MOs were cocultured with FⅫ for 12h.IL-10 was at 3h;the mRNA of MO,including IL-8、IL-10、CCL18、CXCR2 and CCR2,displayed increased expression after FⅫ stimulation.When C1-INH was added,the mRNA expression of related cytokines was decreased.Conclusion：MO stimulated by FⅫ exhibites CD14highCD163highIL-10highCCL18highIL-8high phenotype.FⅫ can induce MO differentiation into a TAM-like macrophage.

关 键 词：凝血因子Ⅻ 单核细胞 肿瘤相关巨噬细胞 卵巢肿瘤 

分 类 号：R711.75[医药卫生—妇产科学]