title: Experimental evidences for hsa-miR-497-5p as a negative regulator of SMAD3 gene expression.
abstract: The SMAD family comprises of transcription factors that function as signal transducers of transforming growth factor (TGFβ) superfamily members. MiRNAs are a class of small noncoding RNAs that may play a major role in post transcriptional regulation of SMAD genes. Here, we intended to investigate if hsa-miR-497-5p is capable of regulating SMAD3 gene expression. Hsa-miR-497-5p was bioinformatically predicted as a candidate regulator of SMAD3 gene expression and then, hsa-miR-497-5p expression status was analyzed in different cell lines using RT-qPCR. Overexpression of hsa-miR-497-5p in HEK293t cells resulted in downregulation of SMAD3 which was detected by RT-qPCR and western analysis. Further, dual luciferase assay results supported direct interaction of hsa-miR-497-5p with 3'-UTR sequences of SMAD3 transcript. Overexpression of hsa-miR-497-5p in HEK293t cells resulted in cell cycle arrest in G0/G1 phase, detected by flow cytometry. Overall, accumulative results indicated that hsa-miR-497-5p by targeting SMAD3 is potentially one of the regulators of the TGFβ signaling pathway.
authors.affiliation: Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Meisam
authors.initials: M
authors.lastname: Jafarzadeh
authors.affiliation: Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran. Electronic address: soltanib@modares.ac.ir.
authors.forename: Bahram M
authors.initials: BM
authors.lastname: Soltani
authors.affiliation: Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Sadat
authors.initials: S
authors.lastname: Dokanehiifard
authors.affiliation: Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Kay
authors.affiliation: Department of Regenerative Medicine, Royan institute, Tehran, Iran.
authors.forename: Nasser
authors.initials: N
authors.lastname: Aghdami
authors.affiliation: Biochemistry Departments, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
mesh_terms: D020413:3' Untranslated Regions; D002453:Cell Cycle; D002460:Cell Line; D015536:Down-Regulation; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D051900:Smad3 Protein / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C559640:MIRN497 microRNA, human; D035683:MicroRNAs; C497790:SMAD3 protein, human; D051900:Smad3 Protein
keywords: Hsa-miR-497-5p; SMAD3; TGFβ signaling pathway