Department of Cardiology, Tianjin Key Laboratory of Ionic‑Molecular Function of Cardiovascular Disease, Tianjin Institute of Cardiology, Second Hospital of Tianjin Medical University, Tianjin 300211, P.R. China.
State Key Laboratory of Medicinal Chemical Biology, School of Medicine, Nankai University, Tianjin 300071, P.R. China.
Mol Med Rep. 2017 Aug;16(2):1307-1313. doi: 10.3892/mmr.2017.6717. Epub 2017 Jun 7.
MiRNA (miR)-128, which is a well‑recognized inhibitor of tumor growth, is involved in the anti-tumor function of dendritic cells (DCs). However, the association between miR‑128 and the DC‑mediated anti‑tumor immunity remains to be elucidated. Murine B16 melanoma cells and C57BL/6 male mice were used to obtain marrow‑derived DCs. DCs were treated with B16 cell suspension. miR‑128 mimic, miR‑128 inhibitor, p38 inhibitor or negative control oligonucleotides were transfected into DCs. After transfection, mRNA and protein expression of p38 in DCs was detected via reverse transcription‑quantitative polymerase chain reaction and western blotting. The present study demonstrated that the miR‑128 abundance in DCs was significantly attenuated by B16 (a melanoma cell line) stimulation and the protein expression level of p38 was increased. Additionally, miR‑128 inhibited the protein expression of p38 in DCs in a dose‑dependent manner, however no significant effect on the p38 mRNA level was observed. Furthermore, miR‑128 mimic or p38 inhibitor decreased the mRNA expression and secretion of interleukin (IL)‑6 and IL‑10 cytokines and increased the level of IL‑12 in DCs, whereas an miR‑128 inhibitor exhibited the opposite effects. These findings suggested that miR‑128 regulated the immune response of DCs via p38‑downstream cytokines. Furthermore, the tumor growth rate, size and weight were markedly decreased and the survival time prolonged, following injection of DCs harboring miR‑128 mimic or p38 inhibitor in C57BL/6 mice bearing B16 melanoma. The results therefore suggest that miR‑128 enhances the anti‑tumor immunity response of DCs via targeting of the p38 mitogen activated protein kinase signaling pathway.
miRNA（miR）-128 是一种公认的肿瘤生长抑制剂，参与树突状细胞（DC）的抗肿瘤功能。然而，miR-128 与 DC 介导的抗肿瘤免疫之间的关联仍有待阐明。使用小鼠 B16 黑色素瘤细胞和 C57BL/6 雄性小鼠获得骨髓来源的 DC。用 B16 细胞悬液处理 DC。将 miR-128 模拟物、miR-128 抑制剂、p38 抑制剂或阴性对照寡核苷酸转染到 DC 中。转染后，通过逆转录-定量聚合酶链反应和 Western blot 检测 DC 中 p38 的 mRNA 和蛋白表达。本研究表明，B16（一种黑色素瘤细胞系）刺激可显著减弱 DC 中的 miR-128 丰度，并增加 p38 蛋白表达水平。此外，miR-128 以剂量依赖的方式抑制 DC 中 p38 的蛋白表达，但对 p38 mRNA 水平没有明显影响。此外，miR-128 模拟物或 p38 抑制剂降低了 DC 中白细胞介素（IL）-6 和 IL-10 细胞因子的 mRNA 表达和分泌，并增加了 IL-12 的水平，而 miR-128 抑制剂则表现出相反的作用。这些发现表明，miR-128 通过 p38 下游细胞因子调节 DC 的免疫反应。此外，在 C57BL/6 小鼠中注射携带 miR-128 模拟物或 p38 抑制剂的 DC 后，B16 黑色素瘤的肿瘤生长速度、大小和重量明显降低，生存时间延长。结果表明，miR-128 通过靶向 p38 丝裂原活化蛋白激酶信号通路增强 DC 的抗肿瘤免疫反应。
