MicroRNA-145 regulates human corneal epithelial differentiation.

<h4>Background</h4>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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sharon Ka-Wai Lee, Yufei Teng, Hoi-Kin Wong, Tsz-Kin Ng, Li Huang, Peng Lei, Kwong-Wai Choy, Yingpeng Liu, Mingzhi Zhang, Dennis Shun-Chiu Lam, Gary Hin-Fai Yam, Chi-Pui Pang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/569dedc6489f4877b06b4141ed2b569b
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:569dedc6489f4877b06b4141ed2b569b
record_format	dspace
spelling	oai:doaj.org-article:569dedc6489f4877b06b4141ed2b569b2021-11-18T06:51:43ZMicroRNA-145 regulates human corneal epithelial differentiation.1932-620310.1371/journal.pone.0021249https://doaj.org/article/569dedc6489f4877b06b4141ed2b569b2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21701675/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>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.<h4>Methodology/principal findings</h4>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 β8 (ITGB8) expression in both human corneal epithelial cells and primary CEPCs.<h4>Conclusion/significance</h4>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.Sharon Ka-Wai LeeYufei TengHoi-Kin WongTsz-Kin NgLi HuangPeng LeiKwong-Wai ChoyYingpeng LiuMingzhi ZhangDennis Shun-Chiu LamGary Hin-Fai YamChi-Pui PangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 6, p e21249 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sharon Ka-Wai Lee Yufei Teng Hoi-Kin Wong Tsz-Kin Ng Li Huang Peng Lei Kwong-Wai Choy Yingpeng Liu Mingzhi Zhang Dennis Shun-Chiu Lam Gary Hin-Fai Yam Chi-Pui Pang MicroRNA-145 regulates human corneal epithelial differentiation.
description	<h4>Background</h4>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.<h4>Methodology/principal findings</h4>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 β8 (ITGB8) expression in both human corneal epithelial cells and primary CEPCs.<h4>Conclusion/significance</h4>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.
format	article
author	Sharon Ka-Wai Lee Yufei Teng Hoi-Kin Wong Tsz-Kin Ng Li Huang Peng Lei Kwong-Wai Choy Yingpeng Liu Mingzhi Zhang Dennis Shun-Chiu Lam Gary Hin-Fai Yam Chi-Pui Pang
author_facet	Sharon Ka-Wai Lee Yufei Teng Hoi-Kin Wong Tsz-Kin Ng Li Huang Peng Lei Kwong-Wai Choy Yingpeng Liu Mingzhi Zhang Dennis Shun-Chiu Lam Gary Hin-Fai Yam Chi-Pui Pang
author_sort	Sharon Ka-Wai Lee
title	MicroRNA-145 regulates human corneal epithelial differentiation.
title_short	MicroRNA-145 regulates human corneal epithelial differentiation.
title_full	MicroRNA-145 regulates human corneal epithelial differentiation.
title_fullStr	MicroRNA-145 regulates human corneal epithelial differentiation.
title_full_unstemmed	MicroRNA-145 regulates human corneal epithelial differentiation.
title_sort	microrna-145 regulates human corneal epithelial differentiation.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/569dedc6489f4877b06b4141ed2b569b
work_keys_str_mv	AT sharonkawailee microrna145regulateshumancornealepithelialdifferentiation AT yufeiteng microrna145regulateshumancornealepithelialdifferentiation AT hoikinwong microrna145regulateshumancornealepithelialdifferentiation AT tszkinng microrna145regulateshumancornealepithelialdifferentiation AT lihuang microrna145regulateshumancornealepithelialdifferentiation AT penglei microrna145regulateshumancornealepithelialdifferentiation AT kwongwaichoy microrna145regulateshumancornealepithelialdifferentiation AT yingpengliu microrna145regulateshumancornealepithelialdifferentiation AT mingzhizhang microrna145regulateshumancornealepithelialdifferentiation AT dennisshunchiulam microrna145regulateshumancornealepithelialdifferentiation AT garyhinfaiyam microrna145regulateshumancornealepithelialdifferentiation AT chipuipang microrna145regulateshumancornealepithelialdifferentiation
_version_	1718424292591403008

MicroRNA-145 regulates human corneal epithelial differentiation.

Ejemplares similares