title: A novel microRNA located in theTrkCgene regulates the Wnt signaling pathway and is differentially expressed in colorectal cancer specimens.
abstract: Tropomyosin receptor kinase C (TrkC) is involved in cell survival, apoptosis, differentiation, and tumorigenesis.TrkCdiverse functions might be attributed to the hypothetical non-coding RNAs embedded within the gene. Using bioinformatics approaches, a novel microRNA namedTrkC-miR2was predicted within theTrkCgene capable of regulating the Wnt pathway. For experimental verification of this microRNA, the predictedTrkC-premir2sequence was overexpressed in SW480 cells, which led to the detection of two matureTrkC-miR2isomiRs, and their endogenous forms were detected in human cell lines as well. Later, an independent promoter was deduced forTrkC-miR2after the treatment of HCT116 cells with 5-azacytidine, which resulted in differential expression ofTrkC-miR2andTrkChost gene. RT-quantitative PCR and luciferase assays indicated that theAPC2gene is targeted byTrkC-miR2, and Wnt signaling is up-regulated. Also, Wnt inhibition by using small molecules along withTrkC-miR2overexpression and TOP/FOP flash assays confirmed the positive effect ofTrkC-miR2on the Wnt pathway. Consistently,TrkC-miR2overexpression promoted SW480 cell survival, which was detected by flow cytometry, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, and crystal violate analysis. RT-qPCR analysis revealed thatTrkC-miR2is significantly up-regulated (∼70 times) in colorectal tumor tissues compared with their normal pairs. Moreover, theTrkC-miR2expression level discriminated grades of tumor malignancies, which was consistent with its endogenous levels in HCT116, HT29, and SW480 colorectal cancer cell lines. Finally, an opposite expression pattern was observed forTrkC-miR2and theAPC2gene in colorectal cancer specimens. In conclusion, here we introduceTrkC-miR2as a novel regulator of Wnt signaling, which might be a candidate oncogenic colorectal cancer biomarker.
authors.affiliation: From the Department of Molecular Genetics, Faculty of Biological Sciences and.
authors.forename: Sadat
authors.initials: S
authors.lastname: Dokanehiifard
authors.affiliation: From the Department of Molecular Genetics, Faculty of Biological Sciences and.
authors.forename: Atena
authors.initials: A
authors.lastname: Yasari
authors.affiliation: From the Department of Molecular Genetics, Faculty of Biological Sciences and.
authors.forename: Hadi
authors.initials: H
authors.lastname: Najafi
authors.affiliation: From the Department of Molecular Genetics, Faculty of Biological Sciences and.
authors.forename: Meisam
authors.initials: M
authors.lastname: Jafarzadeh
authors.affiliation: Department of Nano-Biotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran 111-14115.
authors.forename: Maryam
authors.initials: M
authors.lastname: Nikkhah
authors.affiliation: From the Department of Molecular Genetics, Faculty of Biological Sciences and.
authors.forename: Seyed Javad
authors.initials: SJ
authors.lastname: Mowla
authors.affiliation: From the Department of Molecular Genetics, Faculty of Biological Sciences and soltanib@modares.ac.ir.
authors.forename: Bahram M
authors.initials: BM
authors.lastname: Soltani
mesh_terms: D014408:Biomarkers, Tumor / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D012334:RNA, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D020812:Receptor, trkC; D060449:Wnt Signaling Pathway
chemical_list: C115681:APC2 protein, human; D014408:Biomarkers, Tumor; D003598:Cytoskeletal Proteins; D035683:MicroRNAs; D012334:RNA, Neoplasm; D020812:Receptor, trkC
keywords: Biomarker; Colorectal cancer; Survival; TrkC-miR2; Wnt signaling; annexin; biomarker; cancer; cell cycle; colorectal cancer