Upregulation of miR-665 promotes apoptosis and colitis in inflammatory bowel disease by repressing the endoplasmic reticulum stress components XBP1 and ORMDL3.
Upregulation of miR-665 promotes apoptosis and colitis in inflammatory bowel disease by repressing the endoplasmic reticulum stress components XBP1 and ORMDL3.
复制标题
miR-665的上调通过抑制内质网应激成分XBP1和ORMDL3促进炎症性肠病中的细胞凋亡和结肠炎
DOI：
10.1038/cddis.2017.76
复制
发表时间：
2017-03-23
影响因子：
9
通讯作者：
Chen M
中科院分区：
文献类型：
--
作者：
Li M;Zhang S;Qiu Y;He Y;Chen B;Mao R;Cui Y;Zeng Z;Chen M
MicroRNA 是基因表达的关键转录后调节因子，也是炎症性肠病 (IBD) 病理生理学的关键介质。本研究旨在研究miR-665在IBD进展中的作用。使用实时 PCR 分析来测定 89 个新鲜分离的 IBD 样本和葡聚糖硫酸钠 (DSS) 诱导的结肠粘膜组织中 miR-665 的表达。通过膜联蛋白 V、TUNEL（末端脱氧核苷酸转移酶 dUTP 缺口末端标记）染色、体外集落形成和 DSS 诱导的体内结肠炎小鼠模型检查 miR-665 在诱导细胞凋亡和结肠炎中的作用。此外，进行荧光素酶报告基因测定、蛋白质印迹分析和微核糖核蛋白免疫沉淀以确定miR-665直接抑制XBP1（X-box结合蛋白-1）和ORMDL3的表达。在此，我们的结果表明 miR-665 在活动性结肠炎中显着上调。功能获得和功能丧失研究表明，miR-665 的异位表达在不同炎症刺激下促进细胞凋亡。重要的是，miR-665模拟物的递送得到促进，而antagomiR-665的注射显着削弱了DSS诱导的体内结肠炎。从机制上讲，我们证明 miR-665 通过抑制 XBP1 和 ORMDL3 诱导细胞凋亡。总而言之，我们的研究结果揭示了 ER 应激信号传导的新调节机制，并表明 miR-665 可能是 IBD 治疗的潜在靶点。
MicroRNAs are critical post-transcriptional regulators of gene expression and key mediators of pathophysiology of inflammatory bowel disease (IBD). This study is aimed to study the role of miR-665 in the progression of IBD. Real-time PCR analysis was used to determine miR-665 expression in 89 freshly isolated IBD samples and dextran sulfate sodium (DSS)-induced colonic mucosal tissues. The role of miR-665 in inducing apoptosis and colitis were examined by Annexin V, TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) staining, colony formation in vitro and DSS-induced colitis mice model in vivo. Moreover, luciferase reporter assay, western blot analysis and microribonucleoprotein immunoprecipitation were performed to determine that miR-665 directly repressed XBP1 (X-box-binding protein-1) and ORMDL3 expression. Herein, our results revealed that miR-665 was markedly upregulated in active colitis. Gain-of-function and loss-of-function studies showed that ectopic expression of miR-665 promoted apoptosis under different inflammatory stimuli. Importantly, delivery of miR-665 mimic promoted, while injection of antagomiR-665 markedly impaired DSS-induced colitis in vivo. Mechanistically, we demonstrated that miR-665 induced apoptosis by inhibiting XBP1 and ORMDL3. Taken together, our findings reveal a new regulatory mechanism for ER stress signaling and suggest that miR-665 might be a potential target in IBD therapy.