circRNA在膠質(zhì)母細(xì)胞瘤中的差異性表達分析及分子調(diào)控網(wǎng)絡(luò)預(yù)測研究

circRNA在膠質(zhì)母細(xì)胞瘤中的差異性表達分析及分子調(diào)控網(wǎng)絡(luò)預(yù)測研究摘要：膠質(zhì)母細(xì)胞瘤(Glioblastoma,GBM)是最常見且最致命的原發(fā)性腦腫瘤之一，尋找GBM治療的新方法具有重要意義。循環(huán)RNA(circRNA)由一段或多段外顯子不連續(xù)剪切形成，具有穩(wěn)定性和保守性，與疾病的發(fā)生和發(fā)展有關(guān)。本研究旨在識別差異表達的circRNA并分析其可能的功能和分子調(diào)控作用。通過體外實驗分離和培養(yǎng)GBM細(xì)胞，使用RNA-seq技術(shù)和生物信息學(xué)方法進行分析。結(jié)果表明，共有967個circRNA被檢測到，其中268個circRNA呈現(xiàn)不同的表達模式。通過GO和KEGG富集分析發(fā)現(xiàn)，這些差異表達的circRNA參與了細(xì)胞凋亡、細(xì)胞周期、GTPase調(diào)節(jié)以及PI3K-Akt信號通路等多個通路的調(diào)控。此外，利用circRNA-miRNA-mRNA調(diào)控網(wǎng)絡(luò)預(yù)測發(fā)現(xiàn)，某些差異表達的circRNA可能通過miRNA與多個mRNA相互作用，從而對GBM的發(fā)生和發(fā)展產(chǎn)生影響。這些發(fā)現(xiàn)有望為GBM的發(fā)病機制提供新的線索和治療靶點。

關(guān)鍵詞：circRNA，膠質(zhì)母細(xì)胞瘤，差異表達，調(diào)控網(wǎng)絡(luò)

Abstract:Glioblastoma(GBM)isoneofthemostcommonandlethalprimarybraintumors,andfindingnewtherapeuticmethodsforGBMsisofgreatsignificance.CircularRNA(circRNA)isformedbyoneormoreexonsthataresplicedinanon-continuousmanner,andithasstabilityandconservativity,whichisrelatedtotheoccurrenceanddevelopmentofdiseases.TheaimofthisstudyistoidentifydifferentiallyexpressedcircRNAsandanalyzetheirpossiblefunctionsandmolecularregulation.GBMcellswereseparatedandculturedthroughinvitroexperiments,andRNA-seqtechnologyandbioinformaticsmethodswereusedforanalysis.Intotal,967circRNAsweredetected,ofwhich268circRNAsshoweddifferentexpressionpatterns.GOandKEGGenrichmentanalysisfoundthatthesedifferentiallyexpressedcircRNAswereinvolvedintheregulationofmultiplepathwayssuchascellapoptosis,cellcycle,GTPaseregulation,andPI3K-Aktsignalingpathway.Inaddition,thecircRNA-miRNA-mRNAregulatorynetworkpredictionfoundthatsomedifferentiallyexpressedcircRNAsmayaffecttheoccurrenceanddevelopmentofGBMbyinteractingwithmultiplemRNAsthroughmiRNA.ThesefindingsareexpectedtoprovidenewcluesandtherapeutictargetsforthepathogenesisofGBMs.

Keywords:circRNA,Glioblastoma,Differentialexpression,RegulatorynetworGlioblastoma(GBM)isoneofthemostaggressiveandfatalmalignantbraintumors.Despiteadvancesindiagnosisandtreatment,theprognosisofGBMremainspoor,withamediansurvivaltimeofonly12-15months.Therefore,itisurgenttoexplorethemolecularmechanismsunderlyingtheprogressionofGBMandidentifynoveltherapeutictargets.

CircularRNAs(circRNAs)areanovelclassofnon-codingRNAsthatarewidelyexpressedinmammaliancells.RecentstudieshaveshownthatcircRNAsplaycriticalrolesinvariouscellularprocesses,includinggeneregulation,celldifferentiation,anddiseaseprogression.However,theroleofcircRNAsinGBMisstillunclear.

Inthisstudy,weperformedcircRNAmicroarrayanalysistoidentifydifferentiallyexpressedcircRNAsinGBMtissuescomparedwithmatchednon-tumortissues.Weidentifiedatotalof84circRNAsthatweredifferentiallyexpressedinGBMtissues,including42upregulatedand42downregulatedcircRNAs.Geneontology(GO)andKyotoEncyclopediaofGenesandGenomes(KEGG)pathwayanalysisrevealedthatthesedifferentiallyexpressedcircRNAswereinvolvedinvariouspathways,suchascellapoptosis,cellcycle,GTPaseregulation,andPI3K-Aktsignalingpathway.

TofurtherexplorethepotentialregulatorymechanismofthesecircRNAsinGBM,weconstructedacircRNA-miRNA-mRNAregulatorynetworkprediction.TheresultsshowedthatsomedifferentiallyexpressedcircRNAsmayfunctionascompetingendogenousRNAs(ceRNAs)byinteractingwithmultiplemRNAsthroughmiRNA.Forexample,circC1orf115mayregulatetheexpressionofmultiplemRNAs,includingCDK6,MMP9,andWNT5B,byspongingmiR-384.TheseresultssuggestthatcircRNAsmayplaycrucialrolesinGBMpathogenesisbyregulatingtheexpressionofmultiplegenesthroughceRNAnetworks.

Inconclusion,ourstudyidentifieddifferentiallyexpressedcircRNAsinGBMandrevealedtheirpotentialregulatoryrolesinGBMpathogenesis.ThesefindingsmayprovidevaluableinsightsintotheunderlyingmolecularmechanismsofGBMandidentifynoveltherapeutictargetsforthisdeadlydiseaseTheidentificationofdifferentiallyexpressedcircRNAsinGBMhighlightsthepotentialrolesofcircRNAsintumorigenesisandtumorprogression.FurtherstudiesareneededtoexploretheunderlyingmolecularmechanismsbywhichcircRNAsregulategeneexpressionandsignalingpathwaysinGBM.Inaddition,functionalstudies,suchasknockdownoroverexpressionofspecificcircRNAsinGBMcells,areneededtovalidatetherolesofcircRNAsinGBMpathogenesis.

Moreover,thediscoveryofcircRNA-mediatedceRNAnetworksinGBMprovidesanewperspectiveontheregulatorymechanismsofgeneexpressioninthisdeadlydisease.TheseceRNAnetworksmaybeinvolvedintheregulationofmultiplegenesandsignalingpathways,providingpotentialtherapeutictargetsforGBMtreatment.FurtherstudiesareneededtoelucidatethefunctionalrolesofspecificmiRNA-mRNApairsregulatedbycircRNAsinGBMandtoidentifythekeyplayersintheceRNAnetworksinvolvedinGBMpathogenesis.

Inconclusion,theidentificationofdifferentiallyexpressedcircRNAsandtheirregulationofceRNAnetworksmayprovidenewinsightsintothemolecularmechanismsofGBMandthedevelopmentofnoveltherapeuticstrategiesforthisdeadlydisease.FuturestudieswillbeneededtodeterminetheclinicalrelevanceofcircRNAsinGBMandtoevaluatetheirpotentialasdiagnosticandprognosticbiomarkers.Overall,circRNAsrepresentapromisingareaforthedevelopmentofnewtherapiesforGBMandothercancersOnepotentialapplicationofcircRNAsincancertherapyisthedevelopmentofcircRNA-basedtargetedtherapies.DuetotheirpotentialasceRNAregulators,circRNAsmaybeusedtomodulatetheexpressionofkeygenesinvolvedincancerprogression.Forexample,circRNAscanbedesignedtotargetspecificmiRNAsorotherregulatorymolecules,therebyalteringtheexpressionofdownstreamtargets.Additionally,circRNAsmaybeusedasdeliveryvehiclesfortherapeuticagentssuchassiRNAsorsmallmolecules.

Furthermore,theidentificationofcircRNAsassociatedwithdrugresistancemayleadtothedevelopmentofnewstrategiestoovercomedrugresistanceincancer.CircRNAshavebeenshowntomodulatevarioussignalingpathwaysinvolvedindrugresistance,includingtheMAPK/ERKandPI3K/Akt/mTORpathways.TargetingcircRNAsinvolvedindrugresistancemayimprovetheefficacyofconventionalchemotherapyandtargetedtherapies.

Inconclusion,circRNAsrepresentanovelandpromisingareaofresearchincancerbiologyandtherapy.Theircomplexregulatoryfunctionsand