title: Upregulation of microRNA-574-3p in a human gastric cancer cell line AGS by TGF-β1.
abstract: The mechanisms that regulate miR-574-3p expression in cells remain elusive. In the present study, we used real-time PCR assay to demonstrate TGF-β1-induced miR-574-3p upregulation in AGS cells, which was inhibited by TGF-β receptor I inhibitor SB431542. We used a computer search to identify Smad binding sites upstream of the miR-574-3p precursor sequence. We demonstrated that silencing Smad4, but not Smad2 or Smad3, significantly inhibited the TGF-β1-induced miR-574-3p upregulation in AGS cells. Furthermore, TGF-β1 significantly increased the activity of a dual-luciferase reporter that contains the Smad binding sites upstream of the miR-574 precursor sequence. Silencing Smad4 significantly inhibited the TGF-β1-induced increase in the activity of the reporter in AGS cells. ChIP assay showed that Smad4 directly bound to the promoter of miR-574-3p. MiR-574-3p inhibition was effective in eliminating the inhibition of AGS cell proliferation induced by TGF-β1, suggesting that TGF-β1 inducing upregulation of miR-574-3p is functionally significant.
authors.affiliation: Department of Pathogenobiology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, China.
authors.forename: Renwen
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Pathogenobiology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, China.
authors.forename: Mingqi
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Pathogenobiology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, China.
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Sui
authors.affiliation: Department of Pathogenobiology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Ding
authors.affiliation: Department of Pathogenobiology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, China. Electronic address: yangq@jlu.edu.cn.
authors.forename: Qing
authors.initials: Q
authors.lastname: Yang
mesh_terms: D001483:Base Sequence; D001549:Benzamides / Q000494:pharmacology; D001665:Binding Sites; D045744:Cell Line, Tumor; D004149:Dioxoles / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005753:Gastric Mucosa / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051899:Smad2 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051900:Smad3 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051901:Smad4 Protein / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; C558811:MIRN574 microRNA, human; D035683:MicroRNAs; D009841:Oligonucleotides; D034741:RNA, Small Interfering; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; C497796:SMAD4 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D051901:Smad4 Protein; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D008156:Luciferases
keywords: Mammalian homologue of Drosophila Mad and C. elegans Sma (Smad); MicroRNA-574-3p(miR-574-3p); Proliferation; Transforming growth factor-β1(TGF-β1)