中国肿瘤生物治疗杂志2017，Vol.24Issue(7)：721-726,6.DOI:10.3872/j.issn.1007-385x.2017.07.005
NKG2D配体RAE1ε对乳腺癌细胞4T1衍生MDSC功能的影响
Effects of NKG2D ligand RAE1ε on the function of 4T1 mammary cancer cells-derived MDSC
摘要
Abstract
Objective:To explore effect of BaF3 primary B lymphocyte that expresses retinoic acid early transcript 1ε (RAE1ε),ligand of mouse NKG2D,on function of myeloid-derived suppressor cell (MDSC) originated from mouse.Methods:Based on mouse primary B lymphocyte BaF3 line,a BaF3-RAE1ε cell that expressed RAE1ε and a BaF3-mock control cell with empty plasmid were structured.CD 11 b+Gr-1 +MDSC was produced through introduction of 4T1 trumor model in situ.Spleen MDSC was respectively co-cultured with BaF3-RAE1ε cell and BaF3-mock cell that acted as stimulating cells.Flow cytometry assay was used to detect expressions of CD40,CD80,F4/80,CD11c and content of reactive oxygen species (ROS) in the MDSC.ELISA assay was used to test contents ofIL-10,IL-4 ans IFN-γin supernatant of the co-culture.Concentration of nitric oxide (NO) in the supematant was examed by Griess assay.Magnetic beads were used to separate the MDSC in the co-culture system,activity of arginase was detected after splitting.In addition,the separated MDSC co-culture with splenocyte activated by anti-CD3/CD28 antibody.Proliferation of the activated CD3+CD8+ T cell was examed by CFSC assay.Results:The MDSC of mouse spleen derived from 4T1 in situ tumor model was successfully obtained.Comparing with the BaF3-mock cell,effecting of the BaF3-RAE1ε cell stimulation on amounts of IL-4,IFN-γ,IL-10 and NO secreted by the MDSC was not significant (P＞0.05),and on amounts of CD40,CD80,F4/80,CD1 1c and ROS expressed by the MDSC also not significant (P＞0.05).Stimulation of the BaF3-RAE1ε cell did more increase activity of arginase in the MDSC than that of the BaF3-mock cell obviously (156.166± 1.209 vs 110.135±7.356,P＜0.01),and evidently enhanced inhibitory effect of the MDSC on proliferation of CD8+ T cell.Conclusion:RAE1ε could enhance inhibitory function in vitro of the MDSC derived from 4T1 mouse with tumor.关键词
视黄酸早期转录因子1ε/髓系抑制性细胞/精氨酸酶/乳腺癌4T1细胞Key words
retinoic acid early transcript 1ε/myeloid-derived suppressor cell/arginase/4T1 cell of breast cancer分类
医药卫生引用本文复制引用
钱莉,刘阳,叶枫,刘露,王少卿,贾筱琴,傅奕,龚卫娟,田芳..NKG2D配体RAE1ε对乳腺癌细胞4T1衍生MDSC功能的影响[J].中国肿瘤生物治疗杂志,2017,24(7):721-726,6.基金项目
国家自然科学基金资助项目(No.81373130,No.81001308),江苏省自然科学基金资助项目(No.BK2010315) （No.81373130,No.81001308）
扬州大学大学生创新创业训练计划资助项目.Project supported by the National Natural Science Foundation of China (No.81373130,No.81001308),the Natural Science Foundation of Jiangsu Province (No.BK2010315),and the Science and Technology Innovation Fund for Undergraduates of Yangzhou University （No.81373130,No.81001308）
