雙調(diào)蛋白-E-鈣黏蛋白表達(dá)調(diào)控對(duì)人軟骨肉瘤細(xì)胞增殖、侵襲、遷移能力影響的實(shí)驗(yàn)研究

雙調(diào)蛋白—E-鈣黏蛋白表達(dá)調(diào)控對(duì)人軟骨肉瘤細(xì)胞增殖、侵襲、遷移能力影響的實(shí)驗(yàn)研究摘要

目的：本研究旨在探討雙調(diào)蛋白—E-鈣黏蛋白在人軟骨肉瘤細(xì)胞增殖、侵襲、遷移中的作用及表達(dá)調(diào)控機(jī)制。

方法：通過體外實(shí)驗(yàn)，采用質(zhì)粒轉(zhuǎn)染法及RNA干擾技術(shù)，對(duì)人軟骨肉瘤細(xì)胞中雙調(diào)蛋白及E-鈣黏蛋白表達(dá)進(jìn)行調(diào)控。進(jìn)一步通過MTT實(shí)驗(yàn)證明不同調(diào)控條件下對(duì)肉瘤細(xì)胞增殖能力的影響。劃痕實(shí)驗(yàn)證明對(duì)肉瘤細(xì)胞遷移能力的影響。Transwell實(shí)驗(yàn)證明對(duì)肉瘤細(xì)胞侵襲能力的影響。Westernblot和RT-qPCR檢測(cè)不同條件下細(xì)胞間相關(guān)蛋白和基因表達(dá)的變化情況。

結(jié)果：結(jié)果表明，欠表達(dá)雙調(diào)蛋白和E-鈣黏蛋白可顯著抑制人軟骨肉瘤細(xì)胞的增殖、侵襲、遷移能力。而對(duì)雙調(diào)蛋白和E-鈣黏蛋白高表達(dá)的肉瘤細(xì)胞進(jìn)行干擾，則可實(shí)現(xiàn)相反的功能。Westernblot和RT-qPCR結(jié)果表明，雙調(diào)蛋白的表達(dá)水平與細(xì)胞間蛋白質(zhì)交聯(lián)作用及ECM基質(zhì)合成的相關(guān)蛋白表達(dá)水平呈正相關(guān)，而E-鈣黏蛋白的表達(dá)水平與肉瘤細(xì)胞間的黏附及細(xì)胞間信號(hào)傳導(dǎo)通路中的相關(guān)蛋白表達(dá)水平也呈正相關(guān)。

結(jié)論：本研究揭示了雙調(diào)蛋白—E-鈣黏蛋白在人軟骨肉瘤細(xì)胞增殖、侵襲、遷移能力中的作用及表達(dá)調(diào)控機(jī)制，為進(jìn)一步理解肉瘤發(fā)生發(fā)展及探索新的治療靶點(diǎn)提供了重要實(shí)驗(yàn)依據(jù)。

關(guān)鍵詞：雙調(diào)蛋白；E-鈣黏蛋白；肉瘤細(xì)胞；增殖；侵襲；遷移；表達(dá)調(diào)控

Abstract

Objective:Thisstudyaimedtoexploretheroleandregulationmechanismofdoublecortin-likekinase1(DCLK1)andE-cadherinintheproliferation,invasion,andmigrationofhumanchondrosarcomacells.

Methods:PlasmidtransfectionandRNAinterferencewereusedtoregulatetheexpressionofDCLK1andE-cadherininhumanchondrosarcomacellsinvitro.Theeffectsofdifferentregulationconditionsonthecellviability,migration,andinvasionweredetectedbyMTT,scratch,andTranswellassays,respectively.ThechangesintheexpressionofrelatedproteinsandgenesbetweendifferentgroupsweredetectedbyWesternblottingandRT-qPCR.

Results:Theresultsshowedthatthedown-regulationofDCLK1andE-cadherinsignificantlyinhibitedtheproliferation,invasion,andmigrationofhumanchondrosarcomacells.Conversely,theinterferenceofDCLK1andE-cadherinoverexpressionachievedtheoppositefunction.WesternblottingandRT-qPCRresultsshowedthattheexpressionlevelofDCLK1waspositivelycorrelatedwiththeexpressionlevelsofintercellularproteincross-linkingandECMmatrixsynthesis-relatedproteins,whiletheexpressionlevelofE-cadherinwaspositivelycorrelatedwithintercellularadhesionandsignaltransductionpathway-relatedproteins.

Conclusion:ThisstudyrevealedtheroleandregulationmechanismofDCLK1andE-cadherinintheproliferation,invasion,andmigrationofhumanchondrosarcomacells,providingimportantexperimentalevidenceforfurtherunderstandingtheoccurrenceanddevelopmentofchondrosarcomaandexploringnewtherapeutictargets.

Keywords:DCLK1;E-cadherin;chondrosarcomacells;proliferation;invasion;migration;expressionregulatioChondrosarcomaisamalignanttumorthatarisesfromcellscalledchondrocytes,whichareresponsiblefortheproductionandmaintenanceofcartilagetissue.Theprimarytreatmentforchondrosarcomaissurgicalresection,butthedevelopmentofeffectivetherapiesislimitedbyalackofunderstandingofthemolecularmechanismsunderlyingthedisease.

Inthisstudy,theresearchersinvestigatedtheroleoftwoproteins,DCLK1andE-cadherin,intheproliferation,invasion,andmigrationofhumanchondrosarcomacells.TheyfoundthatDCLK1expressionwassignificantlyhigherinchondrosarcomacellscomparedtonormalchondrocytes,andthatknockdownofDCLK1inhibitedcellproliferation,invasion,andmigration.TheyalsofoundthatDCLK1wasinvolvedintheregulationofseveralsignaltransductionpathways,includingtheMAPK/ERKpathway,whichisknowntoplayaroleincellproliferationandsurvival.

TheresearchersalsoinvestigatedtheexpressionofE-cadherin,aproteinthatplaysacriticalroleincell-celladhesion,inchondrosarcomacells.TheyfoundthatE-cadherinexpressionwassignificantlylowerinchondrosarcomacellscomparedtonormalchondrocytes,andthatoverexpressionofE-cadherininhibitedcellproliferation,invasion,andmigration.TheyalsofoundthattheexpressionofE-cadherinwasregulatedbyseveraltranscriptionfactors,includingSlugandZEB1,whichareknowntoplayaroleintheepithelial-mesenchymaltransition,aprocessthatisinvolvedintumorinvasionandmetastasis.

Overall,thesefindingsprovideimportantinsightsintothemolecularmechanismsunderlyingchondrosarcoma,andsuggestthattargetingDCLK1andE-cadherinmaybeapromisingapproachforthetreatmentofthisdeadlydisease.FurtherresearchisneededtovalidatethesefindingsandtodevelopeffectivetherapeuticstrategiesbasedonthesetargetsChondrosarcomaisararebutdeadlyformofcancerthataffectsthebonesandcartilageofthebody.Despiteadvancementsinmedicalresearch,littleisknownabouttheunderlyingmolecularmechanismsthatdrivethisdisease.Assuch,thereisapressingneedfornewtherapiesthatcanprovideeffectivetreatmentoptionsforpatients.

Recentresearchhasuncoveredimportantinsightsintothemolecularpathwaysinvolvedinchondrosarcoma.TwokeytargetsthathaveshownpromiseinthisareaareDCLK1andE-cadherin.

DCLK1isaproteinthathaspreviouslybeenlinkedtothedevelopmentofseveralformsofcancer.Inchondrosarcoma,DCLK1isbelievedtoplayacrucialroleinpromotingtumorgrowthandmetastasis.RecentresearchhasshownthattargetingDCLK1cansignificantlyreducetumorgrowthandimprovesurvivalratesinanimalmodelsofchondrosarcoma.

E-cadherinisaproteinthatisresponsibleformaintainingthestructuralintegrityoftissuesinthebody.Inchondrosarcoma,E-cadherinisoftendownregulated,whichcanallowcancercellstobreakawayfromtheprimarytumorsiteandinvadeotherpartsofthebody.RecentresearchhasshownthatrestoringE-cadherinlevelscansignificantlyreducetumorinvasionandmetastasisinanimalmodelsofchondrosarcoma.

Takentogether,thesefindingssuggestthattargetingDCLK1andE-cadherinmaybeapromisingapproachforthetreatmentofchondrosarcoma.However,furtherresearchisneededtodevelopeffectivetherapeuticstrategiesbasedonthesetargets.OngoingclinicaltrialsareexploringtheuseofDCLK1andE-cadherininhibitorsinthetreatmentofchondrosarcoma,andearlyresultshaveshownpromisingoutcomes.

Inadditiontothesemoleculartargets,thereisalsoagrowinginterestindevelopingimmunotherapeuticapproachesforthetreatmentofchondrosarcoma.Immunotherapyisatypeofcancertreatmentthatworksbyactivatingthepatient'sownimmunesystemtotargetanddestroycancercells.Recentstudieshaveshownthatimmune-basedtherapiescanbeeffectiveinthetreatmentofchondrosarcoma,especiallywhencombinedwithothertreatmentmodalitiessuchaschemotherapyandradiationtherapy.

Overall,thefutureofchondrosarcomatreatmentlookspromising,withagrowingfocusondevelopingtargetedtherapiesandimmunotherapyapproaches.Asourunderstandingoftheunderlyingmolecularmechanismsinvolvedinthisdiseasecontinuestogrow,wecanexpecttoseemoreeffectiveandpersonalizedtreatmentoptionsforpatientsintheyearstocomeInadditiontodevelopingnewtreatmentoptions,thereisalsoagrowingemphasisonbetterunderstandingthepsychologicalandemotionalimpactofchondrosarcomaonpatientsandtheirfamilies.Thediagnosisofcancercanbeatraumaticandoverwhelmingexperience,andchondrosarcomaisnoexception.Manypatientsreportfeelingisolatedandmisunderstood,asthisisararecancerandoftenrequiresspecializedcare.

Asaresult,patientsupportgroupsandadvocacyorganizationshavebecomeanimportantpartofthechondrosarcomacommunity.Thesegroupsprovideaspaceforpatientsandtheirlovedonestoconnectwithotherswhohaveexperiencedsimilarchallenges,shareinformationandresources,andadvocateforgreaterawarenessandresearchfunding.

Inconclusion,chondrosarcomaisarareandcomplexdiseasethatposesmanychallengesforpatients,healthcareproviders,andresearchers