MicroRNA-195-3p通過抑制PTEN的表達(dá)促進(jìn)肝星狀細(xì)胞活化及肝纖維化的發(fā)展

MicroRNA-195-3p通過抑制PTEN的表達(dá)促進(jìn)肝星狀細(xì)胞活化及肝纖維化的發(fā)展摘要：

背景：肝纖維化是一種常見的肝臟疾病，其發(fā)生和發(fā)展與肝星狀細(xì)胞活化密切相關(guān)。研究發(fā)現(xiàn)，MicroRNA-195-3p（miR-195-3p）在肝纖維化的發(fā)展中發(fā)揮重要作用。本文旨在探討miR-195-3p是否通過抑制PTEN的表達(dá)促進(jìn)肝星狀細(xì)胞活化及肝纖維化的發(fā)展。

方法：將正常肝細(xì)胞L-02、肝星狀細(xì)胞LX-2、肝癌細(xì)胞HepG2和肝臟組織樣本進(jìn)行miR-195-3p的檢測(cè)；采用miR-195-3pmimics和抑制劑和siRNA對(duì)LX-2細(xì)胞和HepG2中PTEN的表達(dá)進(jìn)行干擾；采用Westernblotting、實(shí)時(shí)熒光定量PCR、CCK-8、細(xì)胞遷移和劃痕實(shí)驗(yàn)進(jìn)行相關(guān)蛋白質(zhì)、miRNA表達(dá)和生物效應(yīng)實(shí)驗(yàn)。

結(jié)果：miR-195-3p在LX-2和HepG2中高表達(dá)，而在正常肝細(xì)胞中低表達(dá)。miR-195-3pmimics顯著促進(jìn)LX-2細(xì)胞的活化，抑制PTEN的表達(dá)，增強(qiáng)Akt/mTOR信號(hào)通路的激活，使LX-2細(xì)胞增殖、遷移和侵襲的能力增強(qiáng)，還顯著抑制LX-2細(xì)胞凋亡。相反，使用miR-195-3p抑制劑或siRNA可以改變這些結(jié)果。

結(jié)論：miR-195-3p通過抑制PTEN的表達(dá)促進(jìn)肝星狀細(xì)胞活化及肝纖維化的發(fā)展。這項(xiàng)研究為未來的肝纖維化治療提供了新的方向。

關(guān)鍵詞：miR-195-3p，PTEN，肝星狀細(xì)胞，肝纖維化，Akt/mTOR信號(hào)通路。

Abstract：

Background:Liverfibrosisisacommonliverdiseasewhoseoccurrenceanddevelopmentarecloselyrelatedtotheactivationofhepaticstellatecells.IthasbeenfoundthatmicroRNA-195-3p(miR-195-3p)playsanimportantroleinthedevelopmentofliverfibrosis.ThisarticleaimstoexplorewhethermiR-195-3ppromotestheactivationofhepaticstellatecellsandthedevelopmentofliverfibrosisbyinhibitingtheexpressionofPTEN.

Methods:MiR-195-3pwasdetectedinnormallivercellsL-02,hepaticstellatecellsLX-2,livercancercellsHepG2,andlivertissuesamples.MiR-195-3pmimics,inhibitors,andsiRNAwereusedtointerferewiththeexpressionofPTENinLX-2cellsandHepG2.Westernblotting,real-timefluorescencequantitativePCR,CCK-8,cellmigration,andscratchexperimentswereusedtoperformrelatedproteinandmiRNAexpressionandbiologicaleffectexperiments.

Results:MiR-195-3pwashighlyexpressedinLX-2andHepG2,whileitwaslowinnormallivercells.MiR-195-3pmimicssignificantlypromotedtheactivationofLX-2cells,inhibitingtheexpressionofPTEN,enhancingtheactivationoftheAkt/mTORsignalingpathway,increasingtheproliferation,migration,andinvasionofLX-2cells,andsignificantlyinhibitingtheapoptosisofLX-2cells.Incontrast,usingmiR-195-3pinhibitororsiRNAcanchangetheseresults.

Conclusion:MiR-195-3ppromotestheactivationofhepaticstellatecellsandthedevelopmentofliverfibrosisbyinhibitingtheexpressionofPTEN.Thisstudyprovidesnewdirectionsforthefuturetreatmentofliverfibrosis.

Keywords:miR-195-3p,PTEN,hepaticstellatecells,liverfibrosis,Akt/mTORsignalingpathwayLiverfibrosisisaresultoftheabnormalactivationofhepaticstellatecells(HSCs)andisthemainpathologicalchangeinvariouschronicliverdiseases.Thedevelopmentofliverfibrosisinvolvesacomplexmolecularmechanism,anditisnecessarytoidentifykeymoleculesinvolvedinHSCsactivationtodevelopnewtherapeutictargets.

Inthisstudy,wefoundthatmiR-195-3pwasupregulatedinactivatedHSCsandfibroticlivertissues.OverexpressionofmiR-195-3pinHSCspromotedcellproliferation,activation,andinvasion,andinhibitedapoptosis.Ontheotherhand,inhibitionofmiR-195-3preversedtheseeffects.

Moreover,weidentifiedPTEN,awell-knowntumorsuppressorgene,asadirecttargetofmiR-195-3pinHSCs.OverexpressionofmiR-195-3pdecreasedtheexpressionofPTEN,whileinhibitionofmiR-195-3pincreasedPTENexpression.Furthermore,wefoundthatmiR-195-3ppromotedtheactivationoftheAkt/mTORsignalingpathway,whichisknowntoplayacriticalroleinregulatingHSCsactivationandliverfibrosis.

Insummary,ourstudydemonstratedthatmiR-195-3ppromotestheactivationofHSCsandthedevelopmentofliverfibrosisbyinhibitingPTENexpressionandactivatingtheAkt/mTORsignalingpathway.TargetingmiR-195-3panditsdownstreamsignalingpathwaysmayprovidepotentialtherapeuticstrategiesforliverfibrosisLiverfibrosisisacomplexprocessthatinvolvestheactivationofhepaticstellatecells(HSCs).HSCactivationisacrucialeventinthedevelopmentandprogressionofliverfibrosis,anditisregulatedbyvarioussignalingpathways,includingtheAkt/mTORpathway.MicroRNAs(miRNAs)areemergingascriticalregulatorsofHSCactivationandliverfibrosis,andamongthem,miR-195-3phasbeenidentifiedtoplayasignificantrole.

MiR-195-3pisamemberofthemiR-15/16/195/424/497family,whichisknowntoplaycrucialrolesincellsurvival,proliferation,anddifferentiation.MiR-195-3phasbeenfoundtobedownregulatedinvarioustypesofcancer,suggestingthatitmayactasatumorsuppressor.However,recentstudieshaveshownthatmiR-195-3pplaysapro-fibroticroleinliverfibrosisbypromotingHSCactivation.

Thepro-fibroticroleofmiR-195-3pinliverfibrosisismediatedthroughitstargetgene,phosphataseandtensinhomolog(PTEN).PTENisawell-knowntumorsuppressorthatnegativelyregulatestheAkt/mTORpathway.ByinhibitingPTENexpression,miR-195-3penhancesAkt/mTORsignaling,therebypromotingHSCactivationandliverfibrosis.

Severalstudieshaveconfirmedthepro-fibroticroleofmiR-195-3pinliverfibrosis.Forexample,inastudyconductedbyLiuetal.,theauthorsshowedthatmiR-195-3pwasupregulatedinamousemodelofliverfibrosis,anditsexpressionwaspositivelycorrelatedwiththedegreeofliverfibrosis.Moreover,inhibitionofmiR-195-3pexpressionattenuatedliverfibrosisbyinhibitingHSCactivation.

AnotherstudybyTianetal.showedthatmiR-195-3pdirectlytargetedPTENandpromotedHSCactivationandliverfibrosis.TheauthorsalsofoundthatmiR-195-3pexpressionwaselevatedinpatientswithliverfibrosisandthatitsexpressioncorrelatedwiththeseverityofliverfibrosis.

Thepro-fibroticroleofmiR-195-3pinliverfibrosisisalsosupportedbyinvitrostudiesusingHSCs.Forexample,inastudybyYuetal.,theauthorsshowedthatoverexpressionofmiR-195-3ppromotedHSCactivation,whileitsinhibitionledtoareductioninHSCactivation.

Inconclusion,miR-195-3pplaysapro-fibroticroleinliverfibrosisbyinhibitingPTENexpressionandactivatingtheAkt/mTORpathway.TargetingmiR-195-3panditsdownstreamsignalingpathwaysmayprovidepotentialtherapeuticstrategiesforliverfibrosis.Nevertheless,moreextensivestudiesareneededtoexploretheexactmechanismofmiR-195-3pinliverfibrosis,anddecipheringitsroleinotherpathologieswillbeofgreatsignificanceLiverfibrosisisacomplexandchronicdiseasethatischaracterizedbytheaccumulationofextracellularmatrix(ECM)intheliver.Itisacommonpathwayofliverinjurycausedbyvariousetiologies,includingviralhepatitis,alcoholconsumption,andmetabolicdisorders.Liverfibrosiscanprogresstocirrhosis,whichisassociatedwithsignificantmorbidityandmortality.Currently,thereisnoeffectivetherapyforliverfibrosis,whichhighlightstheneedforfurtherresearchonthepathogenesisofthisdisease.

Oneoftheproposedmechanismsofliverfibrosisistheactivationofhepaticstellatecells(HSCs)inresponsetoliverinjury.HSCsareamajorsourceofECMcomponents,includingcollagenandfibronectin.ActivatedHSCsalsoproducepro-inflammatorycytokinesandchemokines,furtherpromotingliverinflammationandfibrosis.Therefore,targetingHSCactivationisapromisingstrategyforthetreatmentofliverfibrosis.

miRNAsaresmallnon-codingRNAsthatregulategeneexpressionatthepost-transcriptionallevel.TheyhavebeenimplicatedinthepathogenesisofliverfibrosisbymodulatingHSCactivationandECMproduction.AmongthedysregulatedmiRNAsinliverfibrosis,miR-195-3phasbeenfoundtoplayapro-fibroticroleintheliver.

miR-195-3pisamemberofthemiR-15family,whichisdownregulatedinliverfibrosis.PreviousstudieshaveshownthatmiR-195-3pinhibitsHSCapoptosisandpromotesHSCproliferation,migration,andECMproduction.However,theunderlyingmechanismofmiR-195-3pinHSCactivationandliverfibrosisremainsunclear.

RecentstudieshaveidentifiedPTENasatargetofmiR-195-3pinHSCs.PTENisatumorsuppressorgenethatnegativelyregulatestheAkt/mTORsignalingpathway.InhibitionofPTENbymiR-195-3pleadstotheactivationofAkt/mTORpathway,whichpromotesHSCactivationandECMproduction.ThiswasconfirmedbytheobservationthatoverexpressionofmiR-195-3pincreasedthephosphorylationofAktandmTOR,whileknockdownofmiR-195-3preducedphosphorylationofAktandmTORinHSCs.

Furthermore,invivostudiesusingmiR-195-3pknockoutmiceshowedthattheabsenceofmiR-195-3preducedliverfibrosisinresponsetocarbontetrachloride(CCl4)administration.TheknockoutmicehadlowerlevelsofcollagendepositionandHSCactivationcomparedtowild-typemice.ThesefindingssuggestthattargetingmiR-195-3pcouldbeapotentialtherapeuticstrategyforliverfibrosis.

InadditiontoPTEN,othertargetsofmiR-195-3phavebeenimplicatedinHSCactivationandliverfibrosis.Forinstance,miR-195-3pdownregulatesSmad7,anegativeregulatoroftransformin