摘要
目的:利用体外共培养实验,观察结肠癌细胞SW480中上皮特异性转录因子Ese-3表达水平的改变对树突状细胞(dendritic cell,DC)分化成熟的影响。方法:利用慢病毒系统获得过表达Ese-3的结肠癌细胞SW480(SW480 Ese-3)及其对照细胞株(SW480 NC)。利用磁珠分选方式,从人外周血单个核细胞(PBMC)中获得CD14^+细胞,经rh GM-CSF、rhIL-4刺激获得不成熟的DC(iDC)。通过Transwell小室将SW480Ese-3和SW480 NC细胞分别与iDC进行间接共培养。流式细胞术检测共培养后的DC细胞中HLA-DR、CD14、CD83、CD86的表达。ELISA检测细胞培养上清中IL-12 p70的水平。结果 :SW480 Ese-3/DC共培养组DC细胞表面标志物HLA-DR、CD83、CD86水平,较SW480 NC/DC共培养组升高,CD14水平降低;SW480Ese-3/DC共培养组上清中IL-12 p70水平较对照组升高。结论 :在体外共培养实验中,SW480细胞过表达Ese-3可促进DC细胞分化成熟。
Objective：To explore the effect of epithelium-specific Ets-3（Ese-3） in colorectal cancer cells on the differentiation and maturation of dendritic cells（DCs） in vitro co-culture experiment.Methods：lentivirus-mediated biological transfection technique was used to build a stable expression of Ese-3 gene in SW480 cell line（SW480 Ese-3） and its relative controls（SW480 NC）.CD14＋ cells were obtained from human peripheral blood mononuclear cells（PBMC） by magnetic bead sorting,and induced to produce iDC by IL-4 and rhGM-CSF.The iDC were stimulated by TNF-α,and co-cultured with SW480 Ese-3 and its control cell line SW480 NC respectively by Transwell chamber.The expression of HLA-DR,CD14,CD83 and CD86 in co-cultured DC cells was detected by flow cytometry.The level of IL-12p70 in supernatant of cell culture was detected by ELISA.Results：The levels of HLA-DR,CD83 and CD86 on DCs in the SW480 Ese-3/DC co-culture group were higher than those in SW480 NC/DC co-culture group and the level of CD14 was decreased.The level of IL-12p70 in SW480 Ese-3/DC culture supernatant were higher than those in the control group.Conclusion：In vitro co-culture experiments,SW480 cells overexpressing Ese-3 can promote the differentiation and maturation of DC cells.
出处
《现代肿瘤医学》
CAS
2018年第4期506-510,共5页
Journal of Modern Oncology
基金
国家自然科学基金面上项目(编号:81372255)
关键词
Ese-3
树突状细胞
结肠癌
分化成熟
肿瘤免疫逃逸
微环境
Ese - 3 ,dendritic cells, colorectal cancer, differentiation and maturation, tumor immune escape, micro-environment
