摘要
目的研究巨噬细胞在结肠炎相关结肠癌发生过程中的数量及表型变化，并初步鉴定募集巨噬细胞的趋化因子。方法联合应用氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）诱导C57BL／6小鼠产生结肠炎相关结肠癌，得到从结肠炎到结肠癌的3个不同阶段的小鼠模型（分别命名为AD1、AD2和AD3）。取结肠组织制作成单细胞悬液后，应用流式细胞仪检测巨噬细胞的百分比及表型。结合细胞因子表达谱芯片数据，初步筛选募集巨噬细胞的趋化因子，并采用实时荧光定量PCR法进一步验证趋化因子的表达变化。结果AOM和DSS联合作用于C57BIM6小鼠可有效地模拟人类结肠炎相关结肠癌。AD1组、AD2组和AD3组小鼠结肠组织巨噬细胞占CD45＋白细胞的比例分别为（9．93±1．28）％、（15．42±1．15）％和（21．25±0．62）％，均明显高于对照组[（2．39±0．54）％，均P〈0．01]。AD1组、AD2组和AD3组结肠组织巨噬细胞表面CD206的阳性表达率分别为（15．03±1．54）％、（8．11±3．70）％和（9．06±1．16）％，均明显低于对照组[（19．43±7．31）％，均P〈0．01]。AD1组、AD2组和AD3组结肠组织巨噬细胞表面CD86的阳性表达率分别为（13．67±2．28）％、（46．73±6．58）％和（76．90±14．32）％，AD2和AD3组结肠组织巨噬细胞表面CD86的阳性表达率均明显高于对照组[（19．37±9．69）％，P〈0．01]。AD1组、AD2组和AD3组结肠组织巨噬细胞表面MHC Ⅱ的阳性表达率分别为（31．10±2．69）％、（33．93±14．08）％和（29．93±1．41）％，均明显低于对照组[（50．30±6．58）％，均P〈0．01]。结肠组织浸润的巨噬细胞主要表型为CD206-CD86＋MHC Ⅱ-的促炎症表型。结合细胞因子表达谱芯片及实时荧光定量PCR验证结果，募集结肠部位巨噬细胞的趋化因子可能为粒细胞集落刺激因子（G-CSF）。结论巨噬细胞在结肠炎相关
Objective To investigate the changes of quantity and phenotype of macrophages during the progress of colitis-associated carcinogenesis, and to identify the chemokines mediating macrophage recruitment. Methods Colitis-associated cancer was induced by azoxymethane （AOM） combined with dextran sulfate sodium （DSS） in C57BL/6 mice. The three sequential developmental stages of colitis associated cancer in the mice were named AD1, AD2 and AD3, respectively. Colon tissues were collected and digested into single-cell suspension. The percentage and phenotype of macrophages in the colon tissues were determined by fluorescence activated cell sorter （FACS）. Protein array and real-time polymerase chain reaction （PCR） were used to predict potential chemotatic factors of macrophages. Results Colitisassociated cancer was effectively induced in C57BL/6 mice using AOM combined with DSS. The percentage of macrophages was gradually elevated in the AD1, AD2 and AD3 groups [ （ 9.93 ± 1.28 ） %, （ 15.42± 1.15 ） %, （ 21.25 ± 0. 62） % ], respectively, significantly higher than that of the control group [ （ 2.39 ± 0.54）%, P〈0.01 ].The macrophages infiltrating the colonic mucosa exhibited mainly a pro-inflammatory phenotype as CD206-CD86±MHC Ⅱ-. The positive rates of CD206 in the AD1, AD2 and AD3 groups were （ 15.03± 1.54） %, （ 8.11 ±3.70） %, and （ 9.06± 1.16） %, respectively, significantly lower than that of the control group [ （19.43±7.31）%, P〈0.01 ]. The positive rates of CD86 in the AD2 and AD3 groups were （ 46.73±6.58 ） % and （ 76.90 ± 14.32） %, respectively, significantly higher than that of the control group [ （19.37±9.69） %, P〈0.01 ） ] .The positive rates of MHC Ⅱ in the AD1, AD2 and AD3 groups were （31.10± 2.69） %, （ 33.93± 14.08） %, and （ 29.93± 1.41 ） %, respectively, significantly lower than that of the control group [ （50.30±6.58）%, P〈0.011. Protein array analysis and real-time PCR data revealed that G-CSF was the potential
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2016年第3期165-171,共7页
Chinese Journal of Oncology
基金
国家重点基础研究发展计划（973计划）（2014CB542103）
北京市自然科学基金（7144237）
科技北京百名领军人才培养工程（Z13110700513001）
北京市科技新星计划（Z13110700413066）
