胶质瘤细胞miR-153上调对树突状细胞分化成熟及Nrf2表达的影响

目的:探讨胶质瘤细胞miR-153上调对树突状细胞分化成熟及Nrf2表达的影响。方法:体外培养小鼠胶质瘤细胞系GL261，随机分为对照组、miR-153 mimics阴性对照组、miR-153 mimics组，以miR-153 mimics阴性对照、miR-153 mimics分别处理GL261，以实时荧光定量PCR（qRT-PCR）检测各组GL261细胞miR-153、VEGF-A及IL-10 mRNA水平；以蛋白免疫印迹法检测各组GL261细胞VEGF-A、IL-10、Nrf2蛋白表达；以流式细胞仪检测DC2.4细胞表面共刺激分子MHC-II、CD80、CD86、CD40表达水平；将小鼠T淋巴细胞系与上述各组DC2.4细胞共培养，以CCK-8检测T细胞增殖情况。结果:与对照组相比，miR-153 mimics组GL261细胞miR-153、VEGF-A及IL-10 mRNA水平，VEGF-A及IL-10蛋白表达明显降低（P＜0.05）。DC2.4表面共刺激分子MHC-II、CD80、CD86及CD40表达水平，DC2.4细胞Nrf2蛋白表达，T细胞活力明显升高（P＜0.05）；miR-153 mimics阴性对照组细胞各指标无明显变化（P＞0.05）。结论:上调miR-153可抑制胶质瘤细胞分泌免疫抑制性因子VEGF-A、IL-10，促进树突状细胞分化成熟并上调其Nrf2蛋白表达。

Effects of up-regulation of miR-153 in glioma cells on differentiation and maturation of dendritic cells and expression of Nrf2

Objective:To investigate the effect of up-regulation of miR-153 on the differentiation and maturation of dendritic cells and the expression of Nrf2 in glioma cells.Methods:Mouse glioma cell line GL261 was cultured in vitro and randomly divided into control group,miR-153 mimics negative control group and miR-153 mimics group.GL261 was treated with mimics negative control and miR-153 mimics respectively.The levels of miR-153,VEGF-A and IL-10 mRNA in GL261 cells were detected by real-time fluorescence quantitative PCR (qRT-PCR).The expressions of VEGF-A,IL-10 and Nrf2 proteins in GL261 cells were detected by Western blot.The expression levels of the costimulatory molecules MHC-II,CD80,CD86 and CD40 on the surface of DC2.4 were detected by flow cytometry,and T lymphocyte lines of mice were co-cultured with DC2.4 cells of the above groups,and T cell proliferation was detected by CCK-8.Results:Compared with the control group,the levels of miR-153,VEGF-A,IL-10 and the expressions of VEGF-A and IL-10 in GL261 cells in the miR-153 mimics group were significantly lower (P＜0.05).The expression levels of MHC-II,CD80,CD86 and CD40 on DC2.4 surface,expression of Nrf2 protein in DC2.4 cells and T cell viability were significantly increased (P＜0.05).There were no significant changes in cell parameters in the mimics negative control group (P＞0.05).Conclusion:Up-regulation of miR-153 can inhibit the glioma cells secrete immunosuppressive factors such as VEGF-A and IL-10,promote the differentiation and maturation of dendritic cells and up-regulate the expression of Nrf2 protein.