Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pone.0021249
Title: MicroRNA-145 regulates human corneal epithelial differentiation
Authors: Lee S.K.-W.
Teng Y.
Wong H.-K.
Ng T.-K.
Huang L.
Lei P.
Choy K.-W.
Liu Y.
Zhang M.
Lam D.S.-C.
Yam G.H.-F. 
Pang C.-P.
Keywords: beta8 integrin
breast cancer resistance protein
connexin 43
cytokeratin 12
cytokeratin 3
integrin
microRNA
protein p63
unclassified drug
MIRN145 microRNA, human
adult
article
cell culture
cell differentiation
controlled study
cornea epithelium
cornea limbus
down regulation
epithelium cell
female
gene expression
human
human cell
human tissue
RNA sequence
stem cell
upregulation
amnion
cytology
DNA microarray
fluorescent antibody technique
genetics
in situ hybridization
in vitro study
metabolism
physiology
polymerase chain reaction
Western blotting
Amnion
Blotting, Western
Cell Differentiation
Cells, Cultured
Epithelium, Corneal
Fluorescent Antibody Technique
Humans
In Situ Hybridization
Limbus Corneae
MicroRNAs
Oligonucleotide Array Sequence Analysis
Polymerase Chain Reaction
Stem Cells
Issue Date: 2011
Citation: Lee S.K.-W., Teng Y., Wong H.-K., Ng T.-K., Huang L., Lei P., Choy K.-W., Liu Y., Zhang M., Lam D.S.-C., Yam G.H.-F., Pang C.-P. (2011). MicroRNA-145 regulates human corneal epithelial differentiation. PLoS ONE 6 (6) : e21249. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0021249
Rights: Attribution 4.0 International
Abstract: Background: Epigenetic factors, such as microRNAs, are important regulators in the self-renewal and differentiation of stem cells and progenies. Here we investigated the microRNAs expressed in human limbal-peripheral corneal (LPC) epithelia containing corneal epithelial progenitor cells (CEPCs) and early transit amplifying cells, and their role in corneal epithelium. Methodology/Principal Findings: Human LPC epithelia was extracted for small RNAs or dissociated for CEPC culture. By Agilent Human microRNA Microarray V2 platform and GeneSpring GX11.0 analysis, we found differential expression of 18 microRNAs against central corneal (CC) epithelia, which were devoid of CEPCs. Among them, miR-184 was up-regulated in CC epithelia, similar to reported finding. Cluster miR-143/145 was expressed strongly in LPC but weakly in CC epithelia (P = 0.0004, Mann-Whitney U-test). This was validated by quantitative polymerase chain reaction (qPCR). Locked nucleic acid-based in situ hybridization on corneal rim cryosections showed miR-143/145 presence localized to the parabasal cells of limbal epithelium but negligible in basal and superficial epithelia. With holoclone forming ability, CEPCs transfected with lentiviral plasmid containing mature miR-145 sequence gave rise to defective epithelium in organotypic culture and had increased cytokeratin-3/12 and connexin-43 expressions and decreased ABCG2 and p63 compared with cells transfected with scrambled sequences. Global gene expression was analyzed using Agilent Whole Human Genome Oligo Microarray and GeneSpring GX11.0. With a 5-fold difference compared to cells with scrambled sequences, miR-145 up-regulated 324 genes (containing genes for immune response) and down-regulated 277 genes (containing genes for epithelial development and stem cell maintenance). As validated by qPCR and luciferase reporter assay, our results showed miR-145 suppressed integrin ?8 (ITGB8) expression in both human corneal epithelial cells and primary CEPCs. Conclusion/Significance: We found expression of miR-143/145 cluster in human corneal epithelium. Our results also showed that miR-145 regulated the corneal epithelium formation and maintenance of epithelial integrity, via ITGB8 targeting. © 2011 Lee et al.
Source Title: PLoS ONE
URI: https://scholarbank.nus.edu.sg/handle/10635/161793
ISSN: 19326203
DOI: 10.1371/journal.pone.0021249
Rights: Attribution 4.0 International
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