肿瘤相关巨噬细胞对人结肠癌SW620细胞生物学功能的影响  

Influence of Tumor-associated Macrophages on Biological Function of SW620 Cell

作　　者：张银旭[1] 刘晓梅[2] 左腾[1] 刘宇[1] 张俊华[1] 

机构地区：[1]辽宁医学院附属第一医院普通外科,辽宁锦州121001 [2]辽宁医学院附属第一医院肿瘤内科,辽宁锦州121001

出　　处：《中国医学科学院学报》2011年第1期71-75,共5页Acta Academiae Medicinae Sinicae

基　　金：辽宁省教育厅重点实验室项目(LS2010102)~~

摘　　要：目的研究肿瘤相关巨噬细胞(TAM)对人结肠癌SW 620细胞生物学功能的影响。方法采用白细胞介素(IL)-4体外诱导人M2型巨噬细胞,W estern blot检测其标志分子CD68、巨噬细胞甘露糖受体(MMR)和诱导型一氧化氮合酶(iNOS)的表达情况。Transwell非接触式共培养TAM和SW 620细胞,酶联免疫吸附法(ELISA)检测TAM分泌的细胞因子IL-10、IL-12、IL-23和转化生长因子-β(TGF-β)水平;活性凝胶电泳迁移分析法(EMSA)检测SW 620细胞中核因子-κB(NF-κB)活性;四氮甲唑蓝法(XTT)和荧光激活细胞分选术(FACS)双标记法分别检测培养后SW 620细胞的增殖和凋亡情况。结果 IL-4诱导的人M2型巨噬细胞可表达CD68和MMR,不表达iNOS。SW 620与M2型巨噬细胞共培养24、48 h后,培养上清液中M2型巨噬细胞分泌的IL-10和TGF-β水平较培养前明显升高(P均<0.01),IL-12和IL-23水平与培养前差异无统计学意义(P均>0.05)。与M2型巨噬细胞共培养24、48 h后,SW 620的NF-κBDNA结合活性较培养前分别降低了72%和75%(P均<0.01),细胞增殖活性分别下降了48%和59%(P均<0.01);凋亡率分别为6.37%和7.68%,明显高于对照组的0.37%(P均<0.01)。结论 TAM可能通过抑制SW 620细胞的NF-κB活性,抑制SW 620细胞增殖,促进SW 620细胞凋亡。Objective To study the influence of tumor-associated macrophages（TAMs） on the biological function of SW620 cell.Methods Macrophage was induced into M2-type macrophage form with interleukin（IL）-4.CD68,macrophage mannose receptor（MMR）,and inducible nitric oxide synthase（iNOS） were analyzed with Western blot.SW620 was co-cultured with TAMs in the Transwell.Cytokines including IL-10,IL-12,IL-23,and tramsforming growth factor-β（TGF-β） were detected with enzyme-linked immunosorbent assay（ELISA）.The activity of nuclear factor-κB（NF-κB） in SW620 was analyzed with electrophoreticmobility shift assay（EMSA）.The proliferation and apoptosis of SW620 cells after co-cultured with TAM were determined with tetrazolium four nitrogen（XTT） assay and fluorescence activated cell sorting（FACS）,respectively.Results IL-4 induced M2 type macrophage expressed CD68 and MMR instead of iNOS.After co-cultured with SW620 for 24 hours and 48 hours,M2 type macrophage secreted higher levels of IL-10 and TGF-β than the pre-culture level（P0.01）,although IL-12 and IL-23 showed no significant differences（P0.05）.The activity of NF-κB in SW620 decreased by 72% and 75% after 24 hours and 48 hours compared with the pre-culture level,respectively（both P0.01）.The activity of proliferation decreased by 48% and 59% and the apoptotic rates increased by 6.37% and 7.68% and 0.37% after 24 hours and 48 hours（all P0.01） compared with the pre-culture levels.Conclusion TAM may inhibit the proliferation and promote the apoptosis of SW620 by suppressing the activity of NF-κB.

关 键 词：肿瘤相关巨噬细胞 SW620细胞 共培养 增殖 凋亡 

分 类 号：R735.34[医药卫生—肿瘤]