Microrna-221 and Microrna-222 Modulate Differentiation and Maturation of Skeletal Muscle Cells 英文参考文献.docVIP

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Microrna-221 and Microrna-222 Modulate Differentiation and Maturation of Skeletal Muscle Cells 英文参考文献.doc

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Microrna-221 and Microrna-222 Modulate Differentiation and Maturation of Skeletal Muscle Cells 英文参考文献

Microrna-221andMicrorna-222ModulateDifferentiation andMaturationofSkeletalMuscleCells BeatriceCardinali1.,LorianaCastellani1,2.,PasqualeFasanaro3,AnnalisaBasso1,StefanoAlema`1 ,Fabio Martelli4,GermanaFalcone1* 1IstitutodiBiologiaCellulare,ConsiglioNazionaledelleRicerche,Monterotondo,Italy,2DipartimentodiScienzeMotorieedellaSalute,Universita`degliStudidiCassino, Cassino,Italy,3IRCCS-PoliclinicoSanDonato,SanDonatoMilanese,Milan,Italy,4LaboratoriodiPatologiaVascolare,IstitutoDermopaticodell’Immacolata,IRCCS,Rome, Italy Abstract Background:MicroRNAs(miRNAs)areaclassofsmallnon-codingRNAsthathaverecentlyemergedasimportantregulators ofgeneexpression.Theynegativelyregulategeneexpressionpost-transcriptionallybytranslationalrepressionandtarget mRNAdegradation.miRNAshavebeenshowntoplaycrucialrolesinmuscledevelopmentandinregulationofmusclecell proliferationanddifferentiation. Methodology/PrincipalFindings:BycomparingmiRNAexpressionprofilingofproliferatingmyoblastsversusdifferentiated myotubes, a number of modulated miRNAs, not previously implicated in regulation of myogenic differentiation, were identified. Among these, miR-221 and miR-222 were strongly down-regulated upon differentiation of both primary and established myogenic cells. Conversely, miR-221 and miR-222 expression was restored in post-mitotic, terminally differentiatedmyotubessubjectedtoSrctyrosinekinaseactivation.Bytheuseofspecificinhibitorsweprovideevidence thatexpressionofmiR-221andmiR-222isunderthecontroloftheRas-MAPKpathway.Bothinmyoblastsandinmyotubes, levelsofthecellcycleinhibitorp27inverselycorrelatedwithmiR-221andmiR-222expression,andindeedweshowthat p27mRNAisadirecttargetofthesemiRNAsinmyogeniccells.EctopicexpressionofmiR-221andmiR-222inmyoblasts undergoing differentiation induced a delay in withdrawal from the cell cycle and in myogenin expression, followed by inhibition of sarcomeric protein accumulation. When miR-221 and miR-222 were expressed in myotubes undergoing maturation,aprofoundalterationof