Wu Xiongbo, Xia Min, Chen Dayang, Wu Fang, Lv Zhifa, Zhan Qiang, Jiao Yang, Wang Wenjie, Chen Guangxia, An Fangmei
Department of Gastroenterology, Wuxi People's Hospital Affiliated to Nanjing Medical University, 299 Qingyang Road, Wuxi, Jiangsu, 214023, China.
School of Radiation Medicine and Protection and Collaborative Innovation Center of Radiation Medicine of Jiangsu Higher Education Institutions, Medical School of Soochow University, Suzhou, 215123, China.
Tumour Biol. 2016 Nov;37(11):15019-15029. doi: 10.1007/s13277-016-5313-6. Epub 2016 Sep 22.
Altered microRNA (miRNA) expression plays a role in cholangiocarcinoma (CCA) development; thus, detection of blood-circulating miRNAs could be useful as CCA markers. This study profiled serum miRNA levels in patients with primary sclerosing cholangitis (PSC) and CCA and then assessed the role of miR-150-5p in CCA progression in vitro. Three samples were randomly selected from each of 50 sera of healthy controls, 30 PSC sera, and 28 CCA sera with matched bile samples for miRNA microarray profiling. The dysregulated miRNAs were confirmed using qRT-PCR, and miR-150-5p was selected for further in vitro and ex vivo studies. The miRNA microarray identified three dysregulated miRNAs in both CCA and PSC samples, while miR-150-5p level was consistently lower in CCA sera, bile, and tissues than in normal control and PSC sera (P < 0.05). Furthermore, levels of miR-150-5p were associated with serum carbohydrate antigen 19-9 (CA19-9) levels and CCA pathological grade. Bioinformatic Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses showed that miR-150-5p could regulate hand-full gene pathways, including cancer pathway (P < 0.01). However, overexpression of miR-150-5p inhibited proliferation, migration, and invasion capability of CCA cells (P < 0.05). Luciferase reporter assay showed that miR-150-5p bound to an oncogene Ets including gene-1 (ELK1), and Western blot data confirmed that miR-150-5p suppressed ELK1 expression in CCA cell lines. These results suggest that reduced miR-150-5p expression could contribute to CCA development and progression due to uncontrolled ELK1 expression. Thus, further study could evaluate miR-150-5p as a novel target and predictor for CCA prevention and treatment.
微小RNA（miRNA）表达改变在胆管癌（CCA）发展中起作用；因此，检测血液循环中的miRNA作为CCA标志物可能有用。本研究分析了原发性硬化性胆管炎（PSC）和CCA患者血清miRNA水平，然后在体外评估了miR-150-5p在CCA进展中的作用。从50份健康对照血清、30份PSC血清和28份CCA血清中各随机选取3份样本，并匹配胆汁样本进行miRNA微阵列分析。使用qRT-PCR确认失调的miRNA，并选择miR-150-5p进行进一步的体外和体内研究。miRNA微阵列在CCA和PSC样本中均鉴定出三种失调的miRNA，而miR-150-5p在CCA血清、胆汁和组织中的水平始终低于正常对照和PSC血清（P<0.05）。此外，miR-150-5p水平与血清糖类抗原19-9（CA19-9）水平和CCA病理分级相关。生物信息学京都基因与基因组百科全书（KEGG）和基因本体论（GO）分析表明，miR-150-5p可调节多个基因通路，包括癌症通路（P<0.01）。然而，miR-150-5p的过表达抑制了CCA细胞的增殖、迁移和侵袭能力（P<0.05）。荧光素酶报告基因检测表明，miR-150-5p与一种原癌基因Ets包括基因-1（ELK1）结合，蛋白质印迹数据证实miR-150-5p在CCA细胞系中抑制ELK1表达。这些结果表明，由于ELK1表达失控，miR-150-5p表达降低可能有助于CCA的发展和进展。因此，进一步研究可评估miR-150-5p作为CCA预防和治疗的新靶点及预测指标。
