肿瘤相关巨噬细胞对肝癌EZH2表达的影响  

The effect of tumor associated macrophages on EZH2 in HCC

作　　者：陈娟[1] 成炜[1] 刘珊珊[1] 黄丽[1] 刘玉荣[1] 马宁芳[1] 

机构地区：[1]广州医科大学基础学院组织学与胚胎学教研室,广东广州510182

出　　处：《解剖学研究》2016年第5期345-350,共6页Anatomy Research

基　　金：广州市教育局"羊城学者"科研项目(12A016G)

摘　　要：目的研究肝癌微环境中肿瘤相关巨噬细胞对转录因子EZH2表达的影响及意义。方法用佛波豆寇乙酸(PMA)诱导人单核细胞系THP-1细胞分化为M2型巨噬细胞;用M2型巨噬细胞与肝癌BEL-7402 or QGY-7703细胞共培养方法模拟肝癌免疫微环境,未共培养的肝癌BEL-7402或QGY-7703细胞为空白对照,q RT-PCR和Western blot分别从m RNA和蛋白质水平检测两组肝癌细胞EZH2的表达水平;免疫组织化学检测肿瘤相关巨噬细胞标记分子CD163与EZH2的表达并进行相关性分析。结果 q RT-PCR及细胞形态学显示巨噬细胞诱导成功;巨噬细胞与肝癌细胞QGY-7703、BEL-7402共培养后,EZH2的表达明显下调,与对照组相比差异有统计学意义(P<0.05);肝癌组织芯片免疫组织化学显示肝癌癌巢组织中CD163+巨噬细胞数量明显少于癌旁组织(P<0.05),癌巢组织中巨噬细胞CD163表达分值与肝癌细胞EZH2表达分值呈负相关(r=-0.32,P=0.018)。结论巨噬细胞与肝癌细胞共培养可下调肝癌细胞EZH2的表达;肝癌癌巢组织中巨噬细胞CD163表达分值与肝癌细胞EZH2表达分值呈负相关。Objective To investigate the effects of tumor associated macrophages on the expression of transcription factor EZH2 in hepatocellular carcinoma. Methods The differentiated macrophage （M2）was obtained from PMA stimulated THP-1 cells. BEL-7402 or QGY-7703 HCC cell-macrophage co-culture system was applied to imitate the tumor immuno-microenvironment of body, BEL-7402 or QGY-7703 cells cultured alone was used as control. The expression levels of EZH2 in 7402 and 7703 cell lines after co-culturing with M2 macrophages were detected by qRT-PCR and Western-blot method. The expression of EZH2 and TAM bio- marker CD163＋ were tested in HCC tissue microarray slides by immunofluoreseent staining. The correlation between the expression score of EZH2 and CD163＋ was analyzed. Results Macrophages were successfully induced to become M2 judging from the cell mor- phology and the results of qRT-PCR After co-cultured with macrophages, the expression levels of EZH2 in BEL-7402 and QGY-7703 was significantly reduced （P〈0.05）. The results from immunohistochemistry staining showed that the positive numbers of CD163＋ macrophages in tumor adjacent tissues were more than that of tumor tissues, and in the macrophages infiltrated area of tumor tissue, the expression of EZH2 were decreased （r=-0.32, P=0.018）. Conclusion A significant decrease of EZH2 expression was appeared in HCC cells after co-culturing with M2 when compared with that of control, the expression score of CD163 was negatively correlated with that of EZH2 in HCC.

关 键 词：肝癌 EZH2 巨噬细胞 免疫微环境 

分 类 号：R735.7[医药卫生—肿瘤]