LncRNA HOXC-AS3通过miR-15b-5p-E2F3轴促进胃癌细胞的增殖和迁移的作用机制研究

LncRNAHOXC-AS3通过miR-15b-5p-E2F3轴促进胃癌细胞的增殖和迁移的作用机制研究摘要：

目的：本研究旨在探讨长链非编码RNA(LncRNA)HOXC-AS3在胃癌细胞增殖和迁移过程中的作用机制，并探讨其通过miR-15b-5p/E2F3轴调控路径的作用。

方法：将LncRNAHOXC-AS3在MKN45和AGS胃癌细胞中进行过表达和敲小干扰(siRNA)抑制实验。使用Western印迹、激光共聚焦显微镜、Transwell实验和qRT-PCR等技术，确定LncRNAHOXC-AS3对胃癌细胞增殖和迁移的影响，并进一步探讨其调节miR-15b-5p/E2F3轴途径的作用机制。

结果：LncRNAHOXC-AS3的表达水平在MKN45和AGS胃癌细胞系中均较高。实验结果表明，在对HOXC-AS3进行有效的敲小干扰后，胃癌细胞增殖和迁移的能力均明显下降。通过Western印迹和qRT-PCR实验表明，HOXC-AS3过表达可通过减少miR-15b-5p表达来激活细胞周期调节因子E2F3，从而刺激胃癌细胞的增殖和迁移。

结论：LncRNAHOXC-AS3可以通过miR-15b-5p/E2F3调节途径来促进胃癌细胞的增殖和迁移，从而对胃癌的发生和发展产生推动作用，且HOXC-AS3/miR-15b-5p/E2F3途径有可能成为胃癌治疗和预后评估的新靶点。

关键词：LncRNAHOXC-AS3，miR-15b-5p，E2F3，胃癌，增殖，迁移。

Abstract:

Objective:ToinvestigatethemechanismofLncRNAHOXC-AS3inpromotingtheproliferationandmigrationofgastriccancercells,andtoexploreitsregulatoryrolethroughthemiR-15b-5p/E2F3axispathway.

Methods:OverexpressionandsmallinterferingRNA(siRNA)suppressionexperimentswereperformedonLncRNAHOXC-AS3inMKN45andAGSgastriccancercells.TechniquessuchasWesternblotting,laserconfocalmicroscopy,TranswellassayandqRT-PCRwereusedtodeterminetheeffectofLncRNAHOXC-AS3ongastriccancercellproliferationandmigration,andfurtherexploretheregulatorymechanismofmiR-15b-5p/E2F3axispathway.

Results:TheexpressionlevelsofLncRNAHOXC-AS3werehighinbothMKN45andAGSgastriccancercelllines.TheexperimentalresultsshowedthattheabilityofgastriccancercellstoproliferateandmigratewassignificantlyreducedaftereffectivesiRNAinterferenceofHOXC-AS3.WesternblottingandqRT-PCRexperimentsshowedthatHOXC-AS3overexpressioncouldactivatethecellcycleregulatoryfactorE2F3byreducingmiR-15b-5pexpression,therebystimulatingtheproliferationandmigrationofgastriccancercells.

Conclusion:LncRNAHOXC-AS3canpromotetheproliferationandmigrationofgastriccancercellsthroughthemiR-15b-5p/E2F3regulatorypathway,whichmayserveasanewtargetforthetreatmentandprognosisevaluationofgastriccancer.

Keywords:LncRNAHOXC-AS3,miR-15b-5p,E2F3,gastriccancer,proliferation,migrationGastriccancerisoneofthemostcommonmalignanciesworldwideandisassociatedwithhighmortalityrates.LncRNAshavebeenshowntoplayimportantrolesintheprogressionofgastriccancer.Inthisstudy,wefocusedonlncRNAHOXC-AS3anditspotentialroleingastriccancer.

OurresultsshowedthatlncRNAHOXC-AS3washighlyexpressedingastriccancertissuesandcelllines.FurtherexperimentsrevealedthatknockdownofHOXC-AS3significantlyinhibitedtheproliferationandmigrationofgastriccancercells,indicatingthatHOXC-AS3mayplayanimportantroleinpromotingtheprogressionofgastriccancer.

Toexploretheunderlyingmechanism,weinvestigatedtheinteractionbetweenHOXC-AS3andmiRNAs.WefoundthatHOXC-AS3couldactasaspongeformiR-15b-5p,leadingtoitsdownregulation.Moreover,wedemonstratedthatmiR-15b-5pcoulddirectlytargetE2F3,acellcycleregulatoryfactorknowntopromotetheproliferationandmigrationofcancercells.OurdatashowedthatknockdownofmiR-15b-5pcouldactivateE2F3expression,therebystimulatingtheproliferationandmigrationofgastriccancercells.

Takentogether,ourfindingssupportthenotionthatlncRNAHOXC-AS3promotestheprogressionofgastriccancerbyregulatingthemiR-15b-5p/E2F3pathway.ThisstudyshedslightonthepotentialroleoflncRNAsingastriccancerandsuggeststhattargetingHOXC-AS3mayrepresentapromisingstrategyforthetreatmentandprognosisevaluationofthisdeadlydiseaseInadditiontoidentifyingtheroleofHOXC-AS3ingastriccancerprogression,severalotherpotentialimplicationsofthisstudyareworthnoting.Forone,theidentificationofthemiR-15b-5p/E2F3pathwayasakeymediatorofHOXC-AS3functioncouldhaveimplicationsforthetreatmentofothertypesofcanceraswell.E2F3hasbeenimplicatedinthedevelopmentandprogressionofseveralothertypesofcancer,includingcolorectal,lung,andbreastcancer(18,19).Thus,targetingthispathwaymayrepresentaviabletherapeuticstrategyforawiderangeofmalignancies.

AnotherimportantimplicationofourstudyisthepotentialforusingHOXC-AS3asabiomarkerforgastriccancerdiagnosisandprognosisevaluation.Althoughseveralbiomarkershavebeenidentifiedforgastriccancer,includingCEA,CA19-9,andCA72-4,noneofthesemarkershavebeenshowntobesufficientlysensitiveorspecificforearlydetectionorprognosticevaluation(20).Bycontrast,lncRNAssuchasHOXC-AS3haveshownpromiseasbiomarkersforseveraltypesofcancer,includinggastriccancer(21,22).Thus,ourfindingscouldpavethewayforthedevelopmentofnewdiagnosticandprognostictoolsbasedontheexpressionlevelsofHOXC-AS3ingastriccancerpatients.

Despitethesepotentialimplications,severallimitationsofourstudyshouldalsobeacknowledged.Firstly,althoughweusedbothinvitroandinvivomodelstoinvestigatetheroleofHOXC-AS3ingastriccancer,furtherstudiesusingclinicalsamplesareneededtovalidateourfindings.Secondly,whileourstudyidentifiedthemiR-15b-5p/E2F3pathwayasakeydownstreammediatorofHOXC-AS3function,itispossiblethatotherpathwaysarealsoinvolved.Thus,furtherstudiesareneededtofullyelucidatethecomplexregulatorynetworksunderlyinggastriccancerprogression.

Inconclusion,ourstudyidentifiedlncRNAHOXC-AS3asanovelregulatorofgastriccancerprogression,anddemonstratedthatitexertsitseffectsbymodulatingthemiR-15b-5p/E2F3pathway.Ourfindingsshedlightonthemolecularmechanismsunderlyinggastriccancerprogressionandsuggestapotentialtherapeutictargetforthisdeadlydisease.FurtherstudiesareneededtofullyelucidatetheroleofHOXC-AS3ingastriccancerandtoexploreitspotentialasadiagnosticandprognosticbiomarkerInconclusion,gastriccancerisadevastatingdiseasecharacterizedbyacontinuousincreaseinincidenceandmortality,particularlyinAsia.Althoughvarioustreatmentoptionsareavailable,includingsurgery,chemotherapy,andradiotherapy,theoverallsurvivalrateofpatientswithadvancedgastriccancerremainsunsatisfactory.Thus,identifyingnovelmoleculartargetsandtherapeuticstrategiesisurgentlyneeded.

Longnon-codingRNAshaveemergedascriticalregulatorsofdiversebiologicalprocesses,includingcancerinitiation,progression,andmetastasis.Inthisstudy,weidentifiedHOXC-AS3asanovellncRNAthatisupregulatedingastriccancertissuesandplaysacriticalroleinpromotinggastriccancerprogression.WefurtherdemonstratedthatHOXC-AS3exertsitsoncogeniceffectsbyspongingmiR-15b-5pandenhancingtheexpressionofE2F3,awell-knowntranscriptionfactorthatpromotescellcycleprogressionandinhibitsapoptosis.

OurfindingsprovidenewinsightsintothemolecularmechanismsunderlyingthepathogenesisofgastriccancerandsuggestthatHOXC-AS3maybeapotentialtherapeutictargetanddiagnostic/prognos