肿瘤相关巨噬细胞促进尤文肉瘤细胞增殖、侵袭及血管拟态形成  

Tumor-associated macrophages promote the proliferation, invasion and vasculogenic mimicry of Ewing sarcoma cells

作　　者：周新[1] 陈佳骏[1] 肖前仁 王滕羽 张中卒[1] 邵高海[1] ZHOU Xin;CHEN Jiajun;XlAO Qianren;WANG Tengyu;ZHANG Zhongzu;SHAO Gaohai(Department of Orthopedics, Yongchuan Hospital of Chongqing Medical University, Chongqing 402160, China;Department of Orthopedics, First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China)

机构地区：[1]重庆医科大学附属永川医院骨科,重庆402160 [2]南昌大学第一附属医院骨科,江西南昌330006

出　　处：《肿瘤》2018年第4期283-290,共8页Tumor

基　　金：国家自然科学基金资助项目(编号:81502329);重庆医科大学附属永川医院研究生创新基金资助项目(编号:YJSCX201604)~~

摘　　要：目的:探究肿瘤相关巨噬细胞对尤文肉瘤细胞增殖、侵袭及血管拟态形成能力的影响及其可能的分子作用机制。方法:体外诱导人单核细胞U937向M2型巨噬细胞分化,实时荧光定量PCR法检测诱导前后单核细胞中M2型巨噬细胞相关因子CD68、CD163及CD206的表达差异。将诱导后的人单核细胞U937与尤文肉瘤细胞A673及SK-ES-1共培养后,分别采用CCK-8法、Transwell小室侵袭实验及血管拟态形成实验检测尤文肉瘤细胞增殖、侵袭及血管拟态形成能力的变化。蛋白质印迹法检测共培养后尤文肉瘤细胞中信号转导与转录激活因子3(signal transducer and activator of transcription 3,STAT3)及其下游的磷酸化STAT3(phospho-STAT3,p-STAT3)、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)和周期蛋白D2(cyclin D2,CCND2)表达的变化。结果:体外诱导后的人单核细胞U937中CD68、CD163及CD206表达水平明显升高(P值均<0.01),提示单核细胞成功分化为M2型巨噬细胞。将诱导后的M2型U937细胞分别与尤文肉瘤细胞A673及SK-ES-1共培养48 h后,A673及SK-ES-1细胞增殖能力较未共培养的对照组明显增强(P值均<0.05),且穿透基质胶的细胞数较对照组明显增多(P值均<0.05),新生血管分支数也较对照组明显增多(P值均<0.05)。共培养后的尤文肉瘤细胞中,STAT3及其下游p-STAT3、MMP2和CCND2蛋白表达水平均较对照组明显升高(P值均<0.01)。结论:肿瘤相关巨噬细胞与尤文肉瘤细胞共培养能够明显促进尤文肉瘤细胞的增殖、侵袭及血管拟态形成,其作用可能是通过激活尤文肉瘤细胞中STAT3通路来实现的。Objective： To investigate the effects of tumor-associated macrophages on the proliferation, invasion and vasculogenic mimicry of Ewing sarcoma cells, and to explore the involved molecular mechanism.Methods： Human monocytic U937 cells were cultured and induced to differentiate into M2- type macrophages in vitro. The expressions of M2-type macrophages-relative cytokines such as CD68, CD163 and CD206 in U937 cells before and after induction were detected by real- time fluorescent quantitative PCR. Ewing sarcoma A673 and SK-ES-1 cells were co-cultured with the induced U937 cells （macrophages）. Then the effects of co-culture with macrophages on the proliferation, invasion and vasculogenic mimicry of Ewing sarcoma cells were performed using CCK-8, Transwell chamber invasion and matrigel tubule formation assays, respectively. The expressions of signal transducer and activator of transcription 3 （STAT3） and its down- stream phospho-STAT3 （p-STAT3）, matrix metalloproteinase 2 （MMP2） and cyclin D2 （CCND2） in Ewing sarcoma cells co-cultured with macrophages were detected by Western blotting.Results： The expression levels of CD68, CD163 and CD206 in monocytic U937 cells were significantly up-regulated after induction in vitro （all P 〈 0.01）, suggesting that the human monocytic U937 cells were differentiated into M2-type macrophages. After the M2-type macrophages were co-cultured with Ewing sarcoma A673 and SK-ES-1 cells for 48 h, the proliferation ability of A673 and SK-ES-1 cells increased significantly as compared with the un-co-cultured control group （both P 〈 0.05）, the number of A673 and SK-ES-1 cells passing through matrigel increased significantly as compared with the control group （both P 〈 0.05）, the branches of new vascular in co-cultured groups were more than those in control group （both P 〈 0.05）, and the expression levels of STAT3 and its down-stream p-STAT3, CCND2 and MMP2 proteins were up-regulated in A673 and SK-ES-1 cells （all P 〈 0.01）.Conclusion： Co

关 键 词：肉瘤 Ewing 巨噬细胞 细胞增殖 肿瘤侵润 血管拟态形成 

分 类 号：R730.262[医药卫生—肿瘤]