凝血因子Ⅱ可诱导单核细胞和巨噬细胞分化为卵巢癌腹膜内肿瘤相关巨噬细胞  

Thrombin induced MO/MA differentiation into a M2 like subset with TAM characteristics in ovarian cancer

作　　者：张婷[1] 马政文[2] 王颖[3] 汪希鹏[1] 王睿黎[1] 狄文[1] 程忠平[4] 李卫平[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,上海200025 [3]上海交通大学医学院免疫所,上海200025 [4]上海市杨浦区中心医院妇产科,上海200090

出　　处：《中国实用妇科与产科杂志》2009年第4期265-268,共4页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：国家自然基金资助项目(30600672)

摘　　要：目的探讨激活的凝血因子Ⅱ(Ⅱ因子)能否诱导外周血单核细胞分化为卵巢上皮性癌(EOC)腹膜微环境肿瘤相关巨噬细胞。方法分离正常女性外周血单核细胞和巨噬细胞(MO/MA)以及EOC腹水内肿瘤相关巨噬细胞(TAM);凝血因子Ⅱ刺激后,检测MO/MA和TAM表面Ⅱ因子受体PAR1、3、4表达,分析MO/MA在Ⅱ因子或(Ⅱ因子+水蛭素)刺激后CD14、CD68和CD163表达比例,ELISA检测Ⅱ因子刺激后巨噬细胞表达的细胞因子,Real-timePCR检测细胞因子、趋化因子mRNA表达。结果MO/MA及TAM细胞表面上均有Ⅱ因子受体PAR1、3、4表达;Ⅱ因子刺激后,MO/MA高表达CD14[(50.23±4.34)%]及CD163[(3.82±1.93)%],与Ⅱ因子+水蛭素刺激组[CD14(32.37±6.24)%,CD163(0.56±0.28)%]及培养液对照组[CD14(40.06±8.06)%,CD163(1.02±0.38)%]相比差异有统计学意义(P<0.05);ELISA结果示Ⅱ因子刺激后IL-8、IL-10及TGF-β分泌表现出时间依耐性特征,刺激后3h表达量达高峰;Real-timePCR显示IL-8、IL-10、IL-12、CXCR2、CCR2、CCL18mRNA表达上调,CXCR1和TGF-β表达下降。结论激活的凝血因子Ⅱ能诱导外周血MO/MA分化为CD163highIL-10highIL-12highCCL18highIL-8high,具有类似肿瘤相关巨噬细胞特征的M2型巨噬细胞亚型;Ⅱ因子可能参与了EOC腹膜微环境浸润的TAM的分化和极化作用。Objective To investigate whether coagulation factor Ⅱ （thrombin） could induce monocyte/macrophage into M2 like subset with tumor associated macrophages characteristics in ovarian cancer. Methods Thrombin receptors, including PAR1, 3, 4 on MO/MA and TAM were detected. Phenotypes of MO/MA stimulated with thrombin and TAM for 5 days were analyzed using FITC-labeled anti-CD14, -CD68, PE-labeled anti-CD163. MO/MA was incubated with thrombin （1U/mL） with/without hirudin （1U/mL） at 3, 12, and 24h. The cell culture supernatants were collected and cytokines expression was measured by ELISA. Real-time PCR was used to detect mRNA expression. Results Thrombin receptors, including PAR1, 2, 3 were expressed on MO/MA and TAM. MO/MA-thrombin stimulation maintained higher percentage of CD14（50.23 ± 4.34）% and CD163 （3.82 ± 1.93 ）% than that of MO/MA with thrombin ＋ hirudin [ CD14 （32.37 ± 6.24） %, CD163 （0.56 ± 0.28） % ] stimulation or MO/MA cultured with medium [ CD14 （40.06 ± 8. 06） %, CD163 （ 1.02 ± 0.38） % ] （P 〈 0.05）, which present TAM phenotype profile. In ELISA experiment, the IL-8, IL-10 and TGF-β demonstrated time-dependent expression profile, which were all at maximal production at 3 hours point with thrombin stimulation. The mRNA of IL-8, IL-10, IL-12, CXCR2, CCR2 and CCL18 displayed increased expression; in the mean time, relatively lower expression was observed in mRNA expression of CXCR1 and TGF-β1. Conclusion MO/MA treated by thrombin exhibitsd CD163^hish IL-10^high IL-12^high CCL18^hish IL-8^high M2 like phenotype. Thrombin skew MO/MA differentiation into a M2 like subset with TAM characteristics.

关 键 词：凝血因子 巨噬细胞 肿瘤相关巨噬细胞 卵巢肿瘤 

分 类 号：R71[医药卫生—妇产科学]