title: Bioinformatics prediction and experimental validation of microRNA-20a targeting Cyclin D1 in hepatocellular carcinoma.
abstract: Hepatocellular carcinoma is the major form of primary liver cancer, which is the second and sixth leading cause of cancer-related death in men and women, respectively. Extensive research indicates that Wnt/β-catenin signaling pathway, which plays a pivotal role in growth, development, and differentiation of hepatocellular carcinoma, is one of the major signaling pathways that is dysregulated in hepatocellular carcinoma. Cyclin D1 is a proto-oncogene and is one of the major regulators of Wnt signaling pathway, and its overexpression has been detected in various types of cancers including hepatocellular carcinoma. Using several validated bioinformatic databases, we predicted that the microRNAs are capable of targeting 3'-untranslated region of Cyclin D1 messenger RNA. According to the results, miR-20a was selected as the highest ranking microRNA targeting Cyclin D1 messenger RNA. Luciferase assay was recruited to confirm bioinformatic prediction results. Cyclin D1 expression was first assessed by quantitative real-time polymerase chain reaction in HepG2 cell line. Afterward, HepG2 cells were transduced by lentiviruses containing miR-20a. Then, the expression of miR-20a and Cyclin D1 was evaluated. The results of luciferase assay demonstrated targeting of 3'-untranslated region of Cyclin D1 messenger RNA by miR-20a. Furthermore, 238-fold decline in Cyclin D1 expression was observed after lentiviral induction of miR-20a in HepG2 cells. The results highlighted a considerable effect of miRNA-20a induction on the down-regulation of Cyclin D1 gene. Our results suggest that miR-20a can be used as a novel candidate for therapeutic purposes and a biomarker for hepatocellular carcinoma diagnosis.
authors.affiliation: 1 Biotechnology Research Center, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Hamzeh
authors.initials: H
authors.lastname: Karimkhanloo
authors.affiliation: 2 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.|3 Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Samira
authors.initials: S
authors.lastname: Mohammadi-Yeganeh
authors.affiliation: 1 Biotechnology Research Center, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.|4 Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Zeinab
authors.initials: Z
authors.lastname: Ahsani
authors.affiliation: 5 Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Paryan
mesh_terms: D020413:3' Untranslated Regions; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism; D019295:Computational Biology; D019938:Cyclin D1 / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism; D035683:MicroRNAs / Q000378:metabolism*; D000090063:Proto-Oncogene Mas; D015398:Signal Transduction
chemical_list: D020413:3' Untranslated Regions; C530104:CCND1 protein, human; C000715470:MAS1 protein, human; C534451:MIRN20a microRNA, human; D035683:MicroRNAs; D000090063:Proto-Oncogene Mas; D019938:Cyclin D1
keywords: Bioinformatics; Cyclin D1; hepatocellular carcinoma; microRNA