4EBP1-eIF4E-Mcl-1轴在HHT与ATO杀伤微环境中AML细胞中的作用研究

4EBP1-eIF4E-Mcl-1轴在HHT与ATO杀伤微环境中AML细胞中的作用研究摘要：

目的：本研究旨在探究4EBP1-eIF4E-Mcl-1轴在HHT与ATO杀伤微环境中AML细胞中的作用。

方法：本实验选用K562和HL60细胞进行研究，分别采用HHT和ATO处理K562和HL60细胞，检测细胞增殖及细胞凋亡情况。然后通过Westernblot和Real-timePCR等技术手段检测细胞信号通路分子的变化，并分析其表达水平及作用机理。

结果：实验结果显示，HHT和ATO均能够抑制AML细胞的增殖，促进细胞凋亡；处理后，4EBP1蛋白对eIF4E蛋白表达下调，Mcl-1蛋白表达上调。进一步的研究显示，4EBP1敲除和eIF4E敲降不仅能够增强HHT和ATO的杀伤效果，还能减少Mcl-1蛋白的表达。

结论：本研究揭示了4EBP1-eIF4E-Mcl-1轴在HHT和ATO协同杀伤微环境中的重要作用，为AML的治疗提供了新的思路。

关键词：4EBP1-eIF4E-Mcl-1轴；AML；HHT；ATO；微环境；细胞增殖；细胞凋亡；细胞信号通路

Abstract：

Objective:Thisstudyaimedtoexploretheroleof4EBP1-eIF4E-Mcl-1axisinAMLcellsunderHHTandATO-inducedmicroenvironment.

Methods:K562andHL60cellswereusedinthisstudy.HHTandATOwereusedtotreatK562andHL60cells,respectively,andcellproliferationandapoptosisweredetected.ThenthechangesofthecellsignalingpathwaymoleculesweredetectedbyWesternblotandReal-timePCR,andtheirexpressionlevelsandmechanismswereanalyzed.

Results:TheresultsshowedthatHHTandATOcouldinhibittheproliferationofAMLcellsandpromotecellapoptosis.Aftertreatment,theexpressionofeIF4Eproteinwasdownregulatedby4EBP1protein,andtheexpressionofMcl-1proteinwasupregulated.Furtherstudieshaveshownthatboth4EBP1knockoutandeIF4EknockdowncannotonlyenhancethekillingeffectofHHTandATO,butalsoreducetheexpressionofMcl-1protein.

Conclusion:Thisstudyrevealedtheimportantroleofthe4EBP1-eIF4E-Mcl-1axisinthemicroenvironmentofHHTandATOco-cytotoxicity,providinganewideaforthetreatmentofAML.

Keywords:4EBP1-eIF4E-Mcl-1axis;AML;HHT;ATO;microenvironment;cellproliferation;cellapoptosis;cellsignalingpathwayInsummary,AMLisahighlymalignantdiseaseofthehematopoieticsystem,whichseriouslythreatenshumanhealth.Therefore,itisimportanttoexplorenewstrategiesforthetreatmentofAML.Inrecentyears,combinationtherapyhasbecomeapromisingapproachforthetreatmentofAML.HHTandATOaretwocommonlyuseddrugsinAMLtreatment,andtheircombinationhasshownsynergisticcytotoxicityeffectsonleukemiacells.However,theunderlyingmechanismsoftheHHTandATOco-cytotoxicityarenotfullyunderstood.

Inthisstudy,weinvestigatedtheroleofthe4EBP1-eIF4E-Mcl-1axisinthemicroenvironmentofHHTandATOco-cytotoxicity.WefoundthatthecombinationofHHTandATOcaninhibittheproliferationandinduceapoptosisofleukemiacells,andthiseffectismediatedbythe4EBP1-eIF4E-Mcl-1axis.Specifically,wedemonstratedthatknockdownof4EBP1oreIF4EcanenhancethekillingeffectofHHTandATOonleukemiacells,andreducetheexpressionofMcl-1protein,whichisakeyregulatorofcellsurvival.

Ourfindingshighlighttheimportanceofthe4EBP1-eIF4E-Mcl-1axisinthemicroenvironmentofHHTandATOco-cytotoxicity,andsuggestthattargetingthispathwaymaybeapromisingstrategyforthetreatmentofAML.Furtherstudiesareneededtoexplorethemechanismsofactionofthe4EBP1-eIF4E-Mcl-1axisandtodevelopnewdrugsthatcantargetthispathwayInadditiontothepotentialtherapeuticimplications,ourstudyalsoshedslightonthemechanismsthatunderliethesynergisticcytotoxicityofHHTandATO.WefoundthatHHTandATOtargetdistinctpathwaysinleukemiacells,withHHTinhibitingtranslationinitiationandATOinducingoxidativestressandDNAdamage.However,thecombinationofHHTandATOresultsinamuchgreaterdecreaseincellviabilitythaneitherdrugalone,suggestingthattheyareactingsynergistically.

OnepossibleexplanationforthissynergyisthatHHTandATOhavecomplementaryeffectsonthe4EBP1-eIF4E-Mcl-1axis.WhileHHTinhibitseIF4E-mediatedtranslation,ATOinducesthedegradationofMcl-1protein.Hence,thecombinationofHHTandATOmayleadtoadoublehitonthispathway,resultinginamoreprofoundinhibitionofcellsurvival.

AnotherpossibilityisthatHHTandATOhaveadditiveeffectsonotherpathwaysthatregulatecellsurvival.Forinstance,bothdrugshavebeenshowntoactivatethep53pathway,whichplaysacrucialroleinapoptosisandcellcyclearrest.Moreover,ATOhasbeenreportedtoinduceautophagy,aprocessbywhichcellsdegradeandrecycledamagedmoleculesandorganelles.Autophagycanhavebothpro-survivalandpro-deatheffects,dependingonthecontextandtheextentofactivation.Therefore,theinterplaybetweenthep53andautophagypathwaysmaycontributetothecytotoxicityofHHTandATO.

Inconclusion,ourstudyprovidesinsightintothemolecularmechanismsthatunderliethesynergisticcytotoxicityofHHTandATOinAMLcells.Wedemonstratethatthe4EBP1-eIF4E-Mcl-1axisisacriticaltargetofthiscombinationtherapy,andsuggestthatdrugsthatinhibitthispathwaymayhavetherapeuticpotentialinAML.FuturestudiesaimedatexploringthemechanismsofactionofHHTandATO,aswellasidentifyingnewdrugsthatcanenhancetheircytotoxicity,maypavethewayforthedevelopmentofmoreeffectiveandlesstoxictreatmentsforAMLFurtherinvestigationsareneededtodeterminetheoptimaldosingandadministrationscheduleofHHTandATOincombinationtherapyforAML.Additionally,thepotentialtoxicityofthistreatmentneedstobeevaluatedinpreclinicalandclinicalstudies.StudieshaveshownthattheuseofHHTandATOincombinationwithotherdrugs,suchascytarabineordecitabine,mayincreasetheirefficacyintreatingAML.

Moreover,theidentificationofbiomarkersthatcouldpredictresponsetoHHTandATOcombinationtherapywouldbevaluableinselectingtheappropriatepatientsforthistreatment.Thedevelopmentofpersonalizedtherapies,basedonthegeneticandmolecularcharacteristicsofthepatient'scancer,isapromisingapproachtoincreasingtreatmentefficacyandminimizingtoxicity.

Inconclusion,thecombinationofHHTandATOhasshownpromisingresultsinpreclinicalandclinicalstudiesforthetreatmentofAML.Thesynergisticcytotoxicityofthesetwodrugshasbeenattributedtotheirabilitytotargetdifferentpathwaysinvolvedincancercellsurvivalandproliferation.The4EBP1-eIF4E-Mcl-1axishasbeenidentifiedasacriticaltargetofthiscombinationtherapy,andfurt