title: miRNA and mRNA expression profiling identifies members of the miR-200 family as potential regulators of epithelial-mesenchymal transition in pterygium.
abstract: The current study investigates whether microRNA (miRNA) regulators of epithelial-mesenchymal transition (EMT), tissue fibrosis, and angiogenesis are differentially expressed in human primary pterygium. Genome-wide miRNA and mRNA expression profiling of paired pterygium and normal conjunctiva was performed in the context of conventional excision of pterygium with autotransplantation of conjunctiva (n = 8). Quantitative real time polymerase chain reaction (qRT-PCR) was used to validate the expression of key molecules previously detected by microarray. In pterygium, 25 miRNAs and 31 mRNAs were significantly differentially expressed by more than two-fold compared to normal conjunctiva. 14 miRNAs were up-regulated (miR-1246, -486, -451, -3172, -3175, -1308, -1972, -143, -211, -665, -1973, -18a, 143, and -663b), whereas 11 were down-regulated (miR-675, -200b-star, -200a-star, -29b, -200b, -210, -141, -31, -200a, -934, and -375). Unsupervised hierarchical cluster analysis demonstrated that members of the miR-200 family were coexpressed and down-regulated in pterygium. The molecular and cellular functions that were most significant to the miRNA data sets were cellular development, cellular growth and proliferation, and cellular movement. qRT-PCR confirmed the expression of 15 of the 16 genes tested and revealed that miR-429 was down-regulated by more than two-fold in pterygium. The concerted down-regulation of four members from both clusters of the miR-200 family (miR-200a/-200b/-429 and miR-200c/-141), which are known to regulate EMT, and up-regulation of the predicted target and mesenchymal marker fibronectin (FN1), suggest that EMT could potentially play a role in the pathogenesis of pterygium and might constitute promising new targets for therapeutic intervention in pterygium.
authors.affiliation: Department of Ophthalmology, Stavanger University Hospital, Stavanger, Norway; Department of Clinical Medicine, Section of Ophthalmology, University of Bergen, Bergen, Norway.
authors.forename: David H
authors.initials: DH
authors.lastname: Engelsvold
authors.forename: Tor P
authors.initials: TP
authors.lastname: Utheim
authors.forename: Ole K
authors.initials: OK
authors.lastname: Olstad
authors.forename: Pedro
authors.initials: P
authors.lastname: Gonzalez
authors.forename: Jon R
authors.initials: JR
authors.lastname: Eidet
authors.forename: Torstein
authors.initials: T
authors.lastname: Lyberg
authors.forename: Anne-Marie S
authors.initials: AM
authors.lastname: Trøseid
authors.forename: Darlene A
authors.initials: DA
authors.lastname: Dartt
authors.forename: Sten
authors.initials: S
authors.lastname: Raeder
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D064592:Autografts; D049109:Cell Proliferation; D003228:Conjunctiva / Q000637:transplantation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D005353:Fibronectins / Q000235:genetics; D005355:Fibrosis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D011625:Pterygium / Q000235:genetics* / Q000601:surgery; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: C067625:FN1 protein, human; D005353:Fibronectins; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger
keywords: ACTA; AQP1; CCL19; CDH1; COL3A1; COL4A2; CTGF; E-Cadherin; EFEMP1; EGF containing fibulin-like extracellular matrix protein 1; EMT; FDR; FN1; IKB; Ingenuity Knowledge Base; LTBP2; MMP2; NOTCH1; NR4A1; PECAM1; Q-RT-PCR; Quantitative real-time reverse transcription polymerase chain reaction; SNAI; SPARC; TGB2; UTR; VIM; VWF; ZEB; Zinc finger E-box binding homeobox; actin, alpha 2, smooth muscle, aorta; aquaporin 1 (Colton blood group); chemokine (C–C motif) ligand 19; collagen, type III, alpha 1; collagen, type IV, alpha 2; connective tissue growth factor; epithelial–mesenchymal transition; false discovery rate; fibronectin 1; gene microarray; latent transforming growth factor beta binding protein 2; mRNA; matrix metallopeptidase 2; miR-200 family; miRNA; microRNA; notch 1; nuclear receptor subfamily 4, group A, member 1; platelet/endothelial cell adhesion molecule 1; pterygium; secreted protein acidic cysteine-rich; snail homolog (Drosophila); transforming growth factor beta 2; untranslated region; vimentin; von Willebrand factor