IL-6对M1型巨噬细胞向肿瘤相关M2型巨噬细胞转化过程的诱导作用  

Inducing effect of IL-6 on the transformation of M1 macrophages to tumor-associated M2 macrophages

作　　者：王青山[1] 倪虹[1] 魏晓丽[1] 李玉皓[1] 向荣[1] 王悦[1] 

机构地区：[1]天津南开大学医学院免疫学教研室,300071

出　　处：《解放军医学杂志》2009年第8期927-931,共5页Medical Journal of Chinese People's Liberation Army

基　　金：973国家重大科学研究计划资助项目(2007CB914804);863国家高技术研究发展计划项目(2007AA021010);天津市应用基础及前沿技术研究计划项目(09JCYBJC10800)

摘　　要：目的共培养小鼠M1型巨噬细胞RAW264·7与乳腺癌细胞4T1,观察白介素6(IL-6)在M1型巨噬细胞向肿瘤相关M2型巨噬细胞转化过程中的作用。方法实验共分为以下3组:A组(4T1细胞),B组(RAW264·7细胞),C组(RAW264·7与4T1以1∶4比例混合培养)。流式细胞技术检测M2型巨噬细胞比例,随后加入IL-6特异性抑制剂激活素A(ACTA,25μg/ml)或重组IL-6(rIL-6,10μg/ml)后再行检测。RT-PCR检测M1、M2型巨噬细胞功能相关细胞因子及IL-6、IL-6Rα的mRNA表达量。ELISA法检测3组细胞上清中IL-6的含量,随后改变两种细胞的混合比例后再行检测。结果RAW264·7和4T1细胞共培养后,M2型巨噬细胞比例显著增加,添加ACTA后该比例显著下降,而添加rIL-6后该比例增加。与B组比较,C组的M2型巨噬细胞相关细胞因子(CCL22、CCL2、YM1、PDGF-c、HIMP、MMP-9、IL-10)mRNA表达水平明显增高,M1型巨噬细胞相关因子(IL-12、IL-15、IL-18)mRNA表达水平明显降低。IL-6mRNA在A组呈微量表达,在B组无表达,在C组表达明显增高,而IL-6RαmRNA在3组中均呈高表达。与A、B组比较,C组细胞上清液中IL-6含量明显增加(P均<0·05);以不同比例将两种细胞混合后发现RAW264·7细胞数量的改变显著影响了共培养细胞上清中IL-6的水平。结论将4T1和RAW264·7细胞共培养后,前者能促进后者分泌IL-6,并诱导其向肿瘤相关M2型巨噬细胞转化。IL-6在转化过程中发挥重要作用,阻断其作用可以遏制转化。Objective By co-cultivation of murine macrophage cells （RAW264. 7） with breast carcinoma cells （4T1） to investigate the inductive effects of IL-6 on transformation of M1 macrophages to tumor-associated M2 macrophages. Methods Three groups were set up in present study： group A （4T1 cells）, group B （RAW264. 7 cells） and group C （co-cultivation of RAW264. 7 and 4T1 in a ratio of 1：4）. The proportion of M2 macrophages was first detected by flow cytometry, and then the detection was done again after addition of IL-6 specific inhibitor activin A （ACTA; 25μg/ml） or recombinant IL-6 （rIL-6; 10μg/ml）. RT-PCR was used to detect the mRNA expressions of M1- and M2 related cytokines and IL-6, IL6Rα. The content of IL6 was determined by ELISA in the 3 groups of supernatants, and then the determination was done again after changing the mix ratio of 4T1 and RAW264. 7 cells. Results The proportion of M2 macrophages increased significantly after co-cultivation of RAW264. 7 with 4T1 cells. The addition of ACTA resulted in a decrease of the proportion, but an increase in the proportion of M2 macrophages was found after an addition of rIL-6. In comparison with group B, the mRNA expressions of M2-related cytokines （CCL22, CCL2, YM1, PDGF-c, HIMP, MMP-9 and IL-10） of group C were increased, and those of M1 related cytokines （IL-12, IL-15 and IL-18） were decreased obviously. A very weak expression of IL-6 mRNA was detected in group A, but high in group C, while no expression was observed in group B. The mRNA expression of IL-6Rα was found in all the 3 groups. In comparison with group A and B, the coment of IL-6 in the supernatant of group C increased significantly （P〈0. 05）. The amount of RAW264. 7 cells threw a significant influence on the level of IL-6 when co-cultivating the cells with 4T1 in different ratios. Conclusions When co-cultivating 4T1 with RAW264. 7, the former may promote the latter to secrete IL-6, and induce the latter to transform into tumor associated M2 macrophages. I

关 键 词：巨噬细胞 乳腺肿瘤 白细胞介素6 细胞转化 肿瘤 

分 类 号：R730.3[医药卫生—肿瘤]