real-time pcr-based analysis of the human bile micrornaome identifies mir-9 as a potential diagnostic biomarker for biliary tract cancer实时pcr分析人类的胆汁micrornaome标识mir-9作为一个潜在的诊断胆道癌的生物标志物.pdfVIP

Real-Time PCR-Based Analysis of the Human Bile MicroRNAome Identifies miR-9 as a Potential Diagnostic Biomarker for Biliary Tract Cancer 1 1 2 1 1 Kengo Shigehara , Shigeki Yokomuro , Osamu Ishibashi , Yoshiaki Mizuguchi , Yasuo Arima , Yutaka 1 1 1 1 1 2 Kawahigashi , Tomohiro Kanda , Ichiro Akagi , Takashi Tajiri , Hiroshi Yoshida , Toshihiro Takizawa *, Eiji Uchida1* 1 Department of Surgery for Organ Function and Biological Regulation, Nippon Medical School, Tokyo, Japan, 2 Department of Molecular Medicine and Anatomy, Nippon Medical School, Tokyo, Japan Abstract Biliary tract cancer (BTC) is often difficult to diagnose definitively, even through histological examination. MicroRNAs (miRNAs) regulate a variety of physiological processes. In recent years, it has been suggested that profiles for circulating miRNAs, as well as those for tissue miRNAs, have the potential to be used as diagnostic biomarkers for cancer. The aim of this study was to confirm the existence of miRNAs in human bile and to assess their potential as clinical biomarkers for BTC. We sampled bile from patients who underwent biliary drainage for biliary diseases such as BTC and choledocholithiasis. PCR-based miRNA detection and miRNA cloning were performed to identify bile miRNAs. Using high-throughput real-time PCR-based miRNA microarrays, the expression profiles of 667 miRNAs were compared in patients with malignant disease (n = 9) and age-matched patients with the benign disease choledocholithiasis (n = 9). We subsequently characterized bile miRNAs in terms of stability and localization. Through cloning and using PCR methods