小鼠自发乳腺癌模型中肿瘤相关巨噬细胞的表型检测与分析  

Detection and analysis of phenotypes of tumor-associated macrophages in mouse model of spontaneous breast cancer

作　　者：任晓杰[1] 任鹏[2] 罗松[2] 冀全博[2] 许猛[2] 陆宁[2] 王岩[2] 

机构地区：[1]第三军医大学,重庆400038 [2]解放军总医院骨科,北京100853

出　　处：《细胞与分子免疫学杂志》2017年第6期721-725,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81402216)

摘　　要：目的通过检测巨噬细胞极化标志性分子表达水平变化分析自发乳腺癌进程中肿瘤相关巨噬细胞(TAM)的表型转化。方法通过基因型鉴定获得自发乳腺癌模型小鼠,并通过检测其肿瘤总体积变化,将小鼠分为肿瘤早期组及晚期组;通过机械法获得肿瘤组织单细胞悬液并通过流式细胞分选获得不同病程的TAM;利用实时定量PCR检测M1型巨噬细胞白细胞介素1β(IL-1β)、IL-27、IL-6、CD80、CD86和M2型巨噬细胞标志性分子精氨酸酶1(Arg1)、IL-10、白细胞介素4受体α(IL-4Rα)、巨噬细胞甘露糖受体1(Mrc1)、几丁质酶样分子3(Chil3/Ym1)水平;通过对比早期与晚期肿瘤组织巨噬细胞极化标志性分子的表达倾向性,分析TAM在肿瘤不同时期的表型。结果早期肿瘤的TAM高表达部分M1型巨噬细胞标志性分子IL-1β、CD80、CD86,晚期肿瘤的TAM高表达IL-6及部分M2型巨噬细胞极化标志性分子Arg1、IL-10、IL-4Rα。结论肿瘤进程中,TAM经历表型转换过程,靶向TAM的表型转换有可能为乳腺癌提供新的治疗方案。Objective To detect the phenotypic conversion of tumor-associated macrophages（ TAMs） through analyzing the expression levels of the polarization-related genes. Methods We identified the spontaneous breast cancer mice by genotyping and characterized them into the early stage and the advanced stage groups according to their tumor size. Single cell suspension of the tumor tissues were obtained by mechanical methods and TAMs of different stages were sorted by flow cytometry. We measured the mRNA levels of M1 macrophages-related genes IL-1β,IL-27,IL-6,CD80,CD86 and M2macrophages-related genes arginase 1（ Arg1）,IL-10,interleukin 4 receptor α（ IL-4Rα）,macrophage mannose receptor 1（ Mrc1）,chitinase-like 3（ Chil3/Ym1） by real-time quantitative PCR in order to analyze the phenotypic conversion of TAMs during the tumor progression. Results In the early stage,TAMs expressed high levels of M1-related genes,such as IL-1β,CD80,CD86. On the contrary,TAMs of advanced stage expressed high levels of M2-related genes,including Arg1,IL-10,IL-4Rα as well as IL-6. Conclusion Phenotypic conversion of TAMs is present in tumor progression,and targeting the phenotypes of TAMs may provide potential therapies for breast cancer.

关 键 词：乳腺癌 肿瘤相关巨噬细胞(TAM) 极化 

分 类 号：R737.9[医药卫生—肿瘤] R73-35[医药卫生—临床医学]