巨噬细胞在结肠炎相关结肠癌发生过程中的变化
陈永康;袁伟;徐玥;齐军;马洁
【期刊名称】《中华肿瘤杂志》 【年(卷),期】2016(038)003
【摘要】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 Colitis-associated 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 [(23.9± 0.54)%, P<0.01].The macrophages infiltrating the colonic mucosa exhibited
mainly a pro-inflammatory phenotype as CD206-CD86+MHCII-.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 MHCⅡin 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<0.01].Protein array analysis and real-time PCR data revealed that G-CSF was the potential chemokine to recruit macrophages in the AOM-DSS mouse model.Conclusion Macrophages infiltrate increasingly during the carcinogenesis and development of colitis-associated cancer, which mostly express CD206-CD86+MHCII-and might be potentially recruited by G-CSF.%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C57BL／6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MHCⅡ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＋MHCⅡ－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性结肠癌的发生和发展过程中募集增多，大部分表现为CD206－CD86＋MHCⅡ－，其趋化因子可能为G-CSF。 【总页数】7页(165-171)
【关键词】结肠肿瘤;结肠炎相关结肠癌;氧化偶氮甲烷;葡聚糖硫酸钠;巨噬细胞 【作者】陈永康;袁伟;徐玥;齐军;马洁
【作者单位】100021北京协和医学院中国医学科学院肿瘤医院检验科;100021
巨噬细胞在结肠炎相关结肠癌发生过程中的变化
