結(jié)直腸癌中SENP1和RNF4依賴蘇木化調(diào)節(jié)ZNF281的降解摘要

目的：探究結(jié)直腸癌中SENPl和RNF4對(duì)ZNF281的調(diào)節(jié)機(jī)制及其在腫瘤發(fā)生、發(fā)展中的作用。

方法：使用siRNA靶向剪斷SENPl和RNF4基因表達(dá)，通過(guò)Westernblotting和免疫熒光染色檢測(cè)ZNF281蛋白水平以及先后用體外腫瘤細(xì)胞“裸核體”組分、GST-taggedZNF281RIZ域蛋白進(jìn)行invitroSUMOylation實(shí)驗(yàn)，確定了SENPl-RNF4模塊調(diào)節(jié)ZNF281的SUMO修飾和降解的機(jī)制。

結(jié)果：在結(jié)直腸癌組織標(biāo)本中，SENPl和RNF4表達(dá)明顯升高，而ZNF281蛋白表達(dá)水平也隨之升高。實(shí)驗(yàn)證明，SENPl和RNF4可通過(guò)調(diào)節(jié)ZNF281的SUMO修飾而影響其降解。同時(shí)，ZNF281的幾個(gè)保守Lys位點(diǎn)序列是SENPl調(diào)節(jié)SUMOylation的靶點(diǎn)。在體內(nèi)注射SENPl和RNF4與腫瘤相關(guān)的miRNA后，腫瘤細(xì)胞增殖顯著減緩，腫瘤體積也顯著減小。

結(jié)論：SENPl-RNF4調(diào)節(jié)ZNF281的SUMOylation及降解，參與腫瘤細(xì)胞的增殖、轉(zhuǎn)移和耐藥性的調(diào)控，是結(jié)直腸癌治療和預(yù)防的潛在靶點(diǎn)。

關(guān)鍵詞：結(jié)直腸癌、SENPl、RNF4、SUMOylation、ZNF281

Abstract

Objective:ToinvestigatetheregulatorymechanismofSENPlandRNF4onZNF281andtheirroleintheoccurrenceanddevelopmentofcolorectalcancer.

Methods:siRNAwasusedtotargetSENPlandRNF4geneexpression,andtheproteinlevelofZNF281wasdetectedbyWesternblottingandimmunofluorescencestaining.InvitroSUMOylationexperimentswereperformedusingnakednucleiandGST-taggedZNF281RIZdomainproteintodeterminethemechanismbywhichtheSENPl-RNF4moduleregulatesZNF281SUMOylationanddegradation.

Results:Incolorectalcancertissuesamples,theexpressionofSENPlandRNF4wassignificantlyincreased,andtheproteinexpressionlevelofZNF281alsoincreased.TheresultsoftheexperimentshowedthatSENPlandRNF4canaffectthedegradationofZNF281byregulatingitsSUMOylation.Atthesametime,severalconservedLyssitesequencesofZNF281arethetargetsofSENPlregulationofSUMOylation.AfterinjectionofmiRNArelatedtoSENPlandRNF4invivo,theproliferationoftumorcellswassignificantlyreduced,andthetumorvolumewassignificantlyreduced.

Conclusion:SENPl-RNF4regulatesZNF281SUMOylationanddegradation,participatesintheregulationoftumorcellproliferation,migrationanddrugresistance,andisapotentialtargetforthepreventionandtreatmentofcolorectalcancer.

Keywords:colorectalcancer,SENPl,RNF4,SUMOylation,ZNF28Colorectalcancerisacommonmalignanttumorthatseriouslythreatenshumanhealth.Inrecentyears,theroleofproteinmodificationinthedevelopmentofcancerhasattractedmoreandmoreattention.SUMOylationisapost-translationalmodificationprocessthatplaysanimportantroleincellularphysiologicalandpathologicalprocesses,includingDNArepair,transcriptionalregulation,andproteindegradation.

TheSENPl-RNF4pathwayhasemergedasacrucialregulatorofproteinSUMOylationandde-SUMOylation.ZNF281isatranscriptionfactorthatplaysanimportantroleintumorcellproliferation,migration,anddrugresistance.ThestudyfoundthatSENPlpromotesZNF281SUMOylation,whileRNF4promotesthedegradationofSUMOylatedZNF281.TheSENPl-RNF4pathwayregulatesthestabilityandactivityofZNF281,whichinturnaffectstumorcellproliferation,migration,anddrugresistance.

ThroughinvivoexperimentsusingmiRNArelatedtoSENPlandRNF4,thestudydemonstratedthattheproliferationoftumorcellswassignificantlyreduced,andthetumorvolumewassignificantlyreducedafterinjectionofthesemiRNAs.ThisindicatesthattheSENPl-RNF4pathwaymaybeapotentialtargetforthepreventionandtreatmentofcolorectalcancer.

Inconclusion,theSENPl-RNF4pathwayplaysanimportantroleinregulatingZNF281SUMOylationanddegradation,whichaffectstumorcellproliferation,migration,anddrugresistance.TargetingthispathwaymayprovideanewtherapeuticstrategyforcolorectalcancerItisimportanttonotethatwhiletheSENPl-RNF4pathwayshowspromiseasatherapeutictargetforcolorectalcancer,furtherresearchisneededtofullyunderstanditsroleincancerdevelopmentandprogression.Additionally,thedevelopmentoftargetedtherapiesforthispathwaywillrequirecarefulconsiderationofpotentialoff-targeteffectsandtheuseofappropriateanimalmodelsforpreclinicaltesting.

FuturestudiesmayalsofocusonidentifyingotherfactorsthatregulateZNF281SUMOylationanddegradation.Forexample,itispossiblethatothermiRNAsorproteinsmayalsoplayaroleinthisprocess.Identifyingtheseadditionalfactorscouldleadtothedevelopmentofmoreeffectivetargetedtherapiesforcolorectalcancer.

Insummary,thediscoveryoftheSENPl-RNF4pathwayanditsroleinregulatingZNF281SUMOylationanddegradationrepresentsanexcitingnewavenueforthedevelopmentoftargetedtherapiesforcolorectalcancer.ContinuedresearchinthisfieldwillbeessentialforidentifyingnewtherapeutictargetsandimprovingtheoutlookforpatientswiththisdiseaseThereisnodoubtthatcolorectalcancerisasignificanthealthconcernworldwide,andnewtargetedtherapiesareurgentlyneededtoimprovepatientoutcomes.ThediscoveryoftheSENPl-RNF4pathwayrepresentsapromisingnewavenuefordevelopingsuchtherapies,asitprovidesinsightsintotheregulationofZNF281SUMOylationanddegradation,whicharecrucialprocessesinthedevelopmentandprogressionofcolorectalcancer.

However,itisclearthatthereisstillmuchtobelearnedabouttheunderlyingmechanismsofcolorectalcancerandthemanyfactorsthatcontributetoitsdevelopmentandprogression.Futureresearchinthisfieldwillundoubtedlyfocusonidentifyingnewtherapeutictargetsanddevelopingmoreeffectivetreatmentoptionsforpatients.

Oneareaofparticularinterestistheemergingfieldofprecisionmedicine,whichaimstotailortreatmentstoindividualpatientsbasedontheiruniquegeneticprofilesandotherfactors.Byidentifyingthespecificmolecularpathwaysinvolvedincolorectalcancerandothertypesofcancer,researchersmaybeabletodeveloppersonalizedtreatmentsthataremoretargetedandeffectivethancurrenttreatments.

Anotherimportantareaofresearchisthedevelopmentofnewdiagnostictoolsforcolorectalcancer,whichcouldenableearlierdetectionandmoreeffectivetreatmentofthedisease.Thereiscurrentlynodefinitivebiomarkerorscreeningtestforcolorectalcancer,andmoreresearchisneededtoidentifyreliablebiomarkersanddevelopmoreaccurateandefficientscreeningmethods.

Inconclusion,thediscoveryoftheSENPl-RNF4pathwayanditsroleinregulatingZNF281SUMOylationanddegradationrepresentsanimportantstepforwardinthedevelopmentoftargetedtherapiesforcolorectalcancer.However,thereisstillmuchtobelearn