Cell-free miR-17-5p as a diagnostic biomarker for gastric cancer inhibits dendritic cell maturation
Cell-free miR-17-5p as a diagnostic biomarker for gastric cancer inhibits dendritic cell maturation
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无细胞 miR-17-5p 作为胃癌诊断生物标志物抑制树突状细胞成熟
DOI：
10.2147/ott.s197682
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发表时间：
2019-01-01
影响因子：
4
通讯作者：
Jiang, Hui-Qing
中科院分区：
文献类型：
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作者：
Cui, Zi-Jin;Xie, Xiao-Li;Jiang, Hui-Qing
目的胃癌患者存在miRNA表达异常和树突状细胞功能缺陷。然而，癌细胞衍生的oncomiR在GC检测和树突状细胞（DC）成熟中的作用在很大程度上仍然难以捉摸。方法采用深度测序法筛选候选miRNAs（8例胃癌血浆和8例对照血浆; 8例胃癌组织和8例癌旁正常胃组织），并对164例血浆和72例福尔马林固定石蜡包埋的胃癌组织进行PCR验证。用受试者工作特征曲线评价其诊断性能。通过流式细胞术测定暴露于从用Cy 3-miR-17- 5 p预转染的BGC-823细胞获得的条件培养基的DC中的Cy 3荧光信号，并通过共聚焦显微镜观察。在体外用miR-17- 5 p转染DC时，检测DC的功能和表型改变。结果深度测序和RT-PCR证实5种共有miRNAs在胃癌患者血浆和组织中表达上调。无细胞miR-17- 5 p在GC检测中上级其他检测，曲线下面积为0.82，并且与淋巴结转移和总生存率低相关。GC细胞穿梭的miR-17- 5 p可以被递送到未成熟的DC，并且它们通过减少成熟标志物（MHC II、CD 80和CD 86分子）的表达来显著抑制LPS刺激的表型成熟。与表型标志物的这些改变一致，功能实验表明miR-17- 5 p触发了对DCs内吞活性的抑制作用，并降低了肿瘤坏死因子-α和IL-12的分泌，同时增强了IL-10的产生。混合淋巴细胞反应显示，miR-17- 5 p抑制DCs的T细胞刺激效应，有利于调节性T细胞的扩增。结论胃癌细胞来源的miR-17- 5 p是一种潜在的胃癌检测生物标志物。miR-17- 5 p被DC摄取后，通过抑制树突状细胞的成熟而减弱抗肿瘤免疫应答。
Purpose Gastric cancer (GC) patients display aberrant miRNA expression and defective dendritic cell function. However, the role of cancer cell-derived oncomiR in GC detection and dendritic cell (DC) maturation remains largely elusive. Methods Candidate miRNAs were selected by deep sequencing (8 GC plasma samples vs 8 control plasma samples; 8 GC tissues vs 8 adjacent normal gastric tissues) and confirmed by PCR with 164 plasma samples and 72 formalin-fixed paraffin-embedded GC tissue samples. Their diagnostic performance was evaluated by receiver operating characteristic curve. Cy3 fluorescence signals in DCs, exposed to conditioned medium obtained from BGC-823 cells pre-transfected with Cy3-miR-17-5p, were determined by flow cytometry and visualized by confocal microscopy. Functional and phenotypical alterations of DCs were assayed when DCs were transfected with miR-17-5p in vitro. Results Deep sequencing and RT-PCR confirmed that five shared miRNAs were upregulated in plasma and tissue samples of GC patients. Cell-free miR-17-5p was superior to others in GC detection with an area under the curve of 0.82, and correlated with lymphatic metastasis and poor overall survival. GC cell-shuttled miR-17-5p can be delivered to immature DCs, and they significantly inhibited LPS-stimulated phenotypic maturation by diminishing the expression of maturation markers (MHC II, CD80 and CD86 molecules). In line with those alterations in the phenotypic markers, functional experiments demonstrated that miR-17-5p triggered an inhibitory effect on DCs endocytic activity and decreased tumor necrosis factor-α and IL-12 secretion, while enhancing IL-10 production. Mixed lymphocyte reaction showed that miR-17-5p inhibited the T cell stimulating effect of DCs and favored regulatory T cells expansion. Conclusion GC cell-derived miR-17-5p is a potential biomarker for GC detection. Taken up by DCs, miR-17-5p weakened antitumor immune responses via inhibiting the maturation of dendritic cells.