Niu Qinghui, Li Xiaoyu, Xia Di, Jiang Yueping, Tian Zibin, Bian Cheng, Zhang Cuiping, Liu Pei, Zhang Fengjuan, Yang Yuling, Wang Guanglan
Department of Infection, Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
Department of Gastroenterology, Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2094-2100. doi: 10.3892/etm.2017.4770. Epub 2017 Jul 11.
The present study aimed to determine the expression of microRNA (miRNA or miR)-186 in tumor tissues and peripheral blood of patients with pancreatic cancer (PC), as well as its mechanism of regulation. A total of 65 patients with PC who underwent surgery between June 2013 and October 2015 were included. In addition, 59 healthy subjects were recruited as controls. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of mRNA and miRNA. Western blotting and enzyme-linked immunosorbent assay were used to determine protein expression. Bioinformatics was employed for the prediction of the target gene of miR-186, whereas dual luciferase reporter assay was performed to identify whether miR-186 directly bound to YAP1 mRNA. Human pulmonary aortic endothelial cells (HPACs) were transfected with ago-miR-186. YAP1 expression in HPACs was silenced by siRNA. MTT assay was used to evaluate the viability of HPACs. YAP1 mRNA and protein expression levels were elevated in PC. In addition, expression levels of miR-186 in PC were downregulated. miR-186 regulated the expression of YAP1 by binding with the 3'-untranslated region of YAP1. Elevated expression of miR-186 inhibited the proliferation of HPACs by downregulating the expression of YAP1. Decreased expression of YAP1 by siRNA reduced the viability of HPACs. The present study demonstrates that YAP1 is upregulated in the tumor tissues and blood of PC patients, and this may be associated with the downregulation of miR-186. In addition, miR-186 may affect the occurrence and development of PC by controlling the proliferation of PC cells via YAP1.
本研究旨在确定微小RNA（miRNA或miR）-186在胰腺癌（PC）患者肿瘤组织和外周血中的表达及其调控机制。纳入了2013年6月至2015年10月期间接受手术的65例PC患者。此外，招募了59名健康受试者作为对照。采用逆转录-定量聚合酶链反应检测mRNA和miRNA的表达。采用蛋白质印迹法和酶联免疫吸附测定法测定蛋白质表达。运用生物信息学预测miR-186的靶基因，同时进行双荧光素酶报告基因测定以鉴定miR-186是否直接与YAP1 mRNA结合。用ago-miR-186转染人肺动脉内皮细胞（HPACs）。用小干扰RNA（siRNA）沉默HPACs中的YAP1表达。采用MTT法评估HPACs的活力。PC中YAP1 mRNA和蛋白质表达水平升高。此外，PC中miR-186的表达水平下调。miR-186通过与YAP1的3'-非翻译区结合来调控YAP1的表达。miR-186表达升高通过下调YAP1的表达抑制HPACs的增殖。用siRNA降低YAP1的表达会降低HPACs的活力。本研究表明，YAP1在PC患者的肿瘤组织和血液中上调，这可能与miR-186的下调有关。此外，miR-186可能通过YAP1控制PC细胞的增殖来影响PC的发生和发展。
