Profiling of downregulated blood-circulating miR-150-5p as a novel tumor marker for cholangiocarcinoma
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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.
KeywordsCholangiocarcinoma miRNA profiling miR-150-5p Tumor invasion
The authors would like to thank our staff in both Departments for their contributions to this study and Genminix Company for their technical assistance. This study was supported in part by grants from the National Natural Science Foundation of China (no. 81502038 to FA, no. 81302382 to YJ, and no. 31400720 to WW), the Natural Science Foundation of Jiangsu Province (no. BK2012098 to FA), and the Wuxi Municipal Science and Technology Development Planning Funds (no. CSZ00N1304 to FA).
Compliance with ethical standards
This study was approved by the Institutional Review Board (IRB) of Nanjing Medical University. All patients provided written informed consent before participation of this study.
Conflicts of interest
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