MicroRNA-145 Regulates Human Corneal Epithelial Differentiation 英文参考文献.docVIP

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MicroRNA-145 Regulates Human Corneal Epithelial Differentiation 英文参考文献.doc

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MicroRNA-145 Regulates Human Corneal Epithelial Differentiation 英文参考文献

MicroRNA-145RegulatesHumanCornealEpithelial Differentiation SharonKa-WaiLee1.,YufeiTeng1.,Hoi-KinWong2.,Tsz-KinNg1,LiHuang1,PengLei1 ,Kwong-Wai Choy2,YingpengLiu3,MingzhiZhang3,DennisShun-ChiuLam1,GaryHin-FaiYam1*,Chi-PuiPang1* 1DepartmentofOphthalmologyandVisualSciences,TheChineseUniversityofHongKong,HongKong,China,2DepartmentofObstetricsandGynaecology,TheChinese UniversityofHongKong,HongKong,China,3JointShantouInternationalEyeCentre,Shantou,China Abstract Background:Epigeneticfactors,suchasmicroRNAs,areimportantregulatorsintheself-renewalanddifferentiationofstem cells and progenies. Here we investigated the microRNAs expressed in human limbal-peripheral corneal (LPC) epithelia containingcornealepithelialprogenitorcells(CEPCs)andearlytransitamplifyingcells,andtheirroleincornealepithelium. Methodology/Principal Findings: HumanLPCepitheliawasextractedforsmall RNAsordissociatedfor CEPCculture.By AgilentHumanmicroRNAMicroarrayV2platformandGeneSpringGX11.0analysis,wefounddifferentialexpressionof18 microRNAsagainstcentralcorneal(CC)epithelia,whichweredevoidofCEPCs.Amongthem,miR-184wasup-regulatedin 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-basedinsituhybridizationoncornealrimcryosectionsshowedmiR-143/145presencelocalizedtotheparabasalcellsof limbalepitheliumbutnegligibleinbasalandsuperficialepithelia.Withholocloneformingability,CEPCstransfectedwith lentiviralplasmidcontainingmaturemiR-145sequencegaverisetodefectiveepitheliuminorganotypiccultureandhad increased cytokeratin-3/12 and connexin-43 expressions and decreased ABCG2 andp63 compared with cells transfected withscrambled sequences. Globalgene expressionwas analyzed using Agilent WholeHumanGenome Oligo Microarray andGeneSpringGX11.0.Witha5-folddifferencecomparedtocellswithscrambledsequences,miR-145up-regulated324 g