title: A method of utrophin up-regulation through RNAi-mediated knockdown of the transcription factor EN1.
abstract: The aim of this study was to induce up-regulation of the dystrophin-related gene UTRN that encodes the protein utrophin, to determine whether this could compensate for the lack of dystrophin function in Duchenne muscular dystrophy. The human UTRN promoter, which contains two putative binding sites for homeobox protein engrailed-1 (EN1), was analysed. It was found that EN1 binding site 2 in the UTRN gene promoter directly interacted with transcription factor EN1 in vitro. Chromatin immunoprecipitation assays of the EN1-UTRN promoter complex from rhabdomyosarcoma and HeLa cell lines confirmed that endogenous EN1 interacted with this region in vivo. The findings suggest that EN1 directly interacts with the UTRN promoter. Small interfering RNA was used to inhibit EN1 gene expression. Higher utrophin mRNA levels were observed in EN1-inhibited cells compared with controls. The increase in utrophin mRNA in rhabdomyosarcoma cells and HeLa cells may have resulted from inhibition of EN1 expression.
authors.affiliation: Senior Profession College, Department of Medical Genetics, China Medical University, Shenyang, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Wang
authors.forename: D-H
authors.initials: DH
authors.lastname: Cao
authors.forename: C-L
authors.initials: CL
authors.lastname: Jin
authors.forename: C-K
authors.initials: CK
authors.lastname: Lin
authors.forename: H-W
authors.initials: HW
authors.lastname: Ma
authors.forename: Y-Y
authors.initials: YY
authors.lastname: Wu
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D016189:Dystrophin / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D020868:Gene Silencing / Q000187:drug effects*; D018398:Homeodomain Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000187:drug effects*; D012333:RNA, Messenger / Q000032:analysis; D034741:RNA, Small Interfering / Q000378:metabolism / Q000494:pharmacology*; D015533:Transcriptional Activation; D015854:Up-Regulation; D049411:Utrophin / Q000235:genetics / Q000378:metabolism
chemical_list: D016189:Dystrophin; C513076:EN1 protein, human; D018398:Homeodomain Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C483599:UTRN protein, human; D049411:Utrophin