title: MicroRNA-205 targets tight junction-related proteins during urothelial cellular differentiation.
abstract: The mammalian bladder urothelium classified as basal, intermediate, and terminally differentiated umbrella cells offers one of the most effective permeability barrier functions known to exist in nature because of the formation of apical uroplakin plaques and tight junctions. To improve our understanding of urothelial differentiation, we analyzed the microRNA (miRNA) expression profiles of mouse urinary tissues and by TaqMan miRNA analysis of microdissected urothelial layers and in situ miRNA-specific hybridization to determine the dependence of these miRNAs on the differentiation stage. Our in situ hybridization studies revealed that miR-205 was enriched in the undifferentiated basal and intermediate cell layers. We then used a quantitative proteomics approach to identify miR-205 target genes in primary cultured urothelial cells subjected to antagomir-mediated knockdown of specific miRNAs. Twenty-four genes were reproducibly regulated by miR-205; eleven of them were annotated as cell junction- and tight junction-related molecules. Western blot analysis demonstrated that antagomir-induced silencing of miR-205 in primary cultured urothelial cells elevated the expression levels of Tjp1, Cgnl1, and Cdc42. Ectopic expression of miR-205 in MDCK cells inhibited the expression of tight junction proteins and the formation of tight junctions. miR-205- knockdown urothelial cells showed alterations in keratin synthesis and increases of uroplakin Ia and Ib, which are the urothelial differentiation products. These results suggest that miR-205 may contribute a role in regulation of urothelial differentiation by modulating the expression of tight junction-related molecules.
authors.affiliation: From the ‡Molecular Medicine Research Center, Chang Gung University, Taoyuan 33302, Taiwan; pjc@mail.cgu.edu.tw.
authors.forename: Pei-Jung Katy
authors.initials: PJ
authors.lastname: Chung
authors.affiliation: §Medical Research and Development, Chang Gung Memorial Hospital, Taoyuan 33375, Taiwan;
authors.forename: Lang-Ming
authors.initials: LM
authors.lastname: Chi
authors.affiliation: ¶Department of Urology, Chang Gung Memorial Hospital, Taoyuan 33375, Taiwan;
authors.forename: Chien-Lun
authors.initials: CL
authors.lastname: Chen
authors.affiliation: ‖Department of Microbiology and Immunology, Chung Shan Medical University, Taichung 40201, Taiwan;
authors.forename: Chih-Lung
authors.initials: CL
authors.lastname: Liang
authors.affiliation: From the ‡Molecular Medicine Research Center, Chang Gung University, Taoyuan 33302, Taiwan;
authors.forename: Chung-Tzu
authors.initials: CT
authors.lastname: Lin
authors.affiliation: From the ‡Molecular Medicine Research Center, Chang Gung University, Taoyuan 33302, Taiwan;
authors.forename: Yu-Xun
authors.initials: YX
authors.lastname: Chang
authors.affiliation: **Department of Information Management, Chang Gung University, Taoyuan 33302, Taiwan; and.
authors.forename: Chun-Hsien
authors.initials: CH
authors.lastname: Chen
authors.affiliation: From the ‡Molecular Medicine Research Center, Chang Gung University, Taoyuan 33302, Taiwan; ‡‡Graduate Institute of Biomedical Science, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan.
authors.forename: Yu-Sun
authors.initials: YS
authors.lastname: Chang
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D008813:Mice, Inbred ICR; D035683:MicroRNAs / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D040901:Proteomics; D012333:RNA, Messenger / Q000378:metabolism; D062725:Tight Junction Proteins / Q000235:genetics / Q000378:metabolism*; D019108:Tight Junctions / Q000378:metabolism; D019459:Urothelium / Q000166:cytology / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D062725:Tight Junction Proteins