高迁移率族蛋白b1诱导大肠癌细胞上皮-间质转化与肿瘤血管新生的机制研究-内科学(消化内科)专业论文 word格式.docx

和2G7mAb）以及其受体的抗体（RAGEAb和TLR4Ab）均被用于处理内皮细胞。而且，我们还采用癌-内皮细胞共培养体系来模拟肿瘤微环境。结果：ds-HMGB1通过受体TLR4促使VEGF-A的分泌，at-HMGB1则通过受体RAGE增强内皮细胞迁移。在癌-内皮细胞共培养体系中，抑制HMGB1及其受体能够减弱HMGB1诱导内皮细胞成血管的能力。结论：HMGB1通过不同氧化-还原状态促使内皮细胞迁移与VEGF-A表达，从而调控肿瘤成血管的各个步骤。关键词：大肠癌，高迁移率族蛋白B1，上皮-间质转化，肿瘤血管新生AbstractPartITheMechanismsofHMGB1inEpithelial-MesenchymalTransitionof ColorectalCarcinomaAim:TheaimofthisstudyistodeterminewhetherandthroughwhichmechanismHMGB1(high-mobilitygroupbox1)inducesEMT(epithelial-to-mesenchymaltransition)incolorectalcarcinoma.Methods:WeevaluatedtheinfluenceofHMGB1anditsreceptorsonEMTmarkers ofcancercells via immunofluorescence,PCRandWesternblot techniques. Furthermore,Matrigel-coatedchamberandscratchwoundassayswereappliedto observetheroleofHMGB1anditsreceptorsoncellinvasion.Also,throughPCRand Westernblottechniques,itwasinvestigatedwhichmatrixmetalloproteinases(MMPs) andsignalingpathwaysinvolvingHMGB1-inducedEMT.Results:ThetreatmentofcancercellswithHMGB1inducedalterationsinthephenotypesconsistentwithEMTand enhancedcellmigrationthrougha process mediatedbythereceptorforadvancedglycationend-products(RAGE).Thelevelsof Snailandphospho-NF-κBwereupregulatedduringtheHMGB1-inducedEMT.In addition,HMGB1increasedtheexpressionofMMP-7butnotthatofMMP-9,and thiseffect wasalsoregulatedbySnail/NF-κBsignalling.Conclusion:HMGB1actsasapotentdriverofcancerEMTthroughtheRAGE/Snail/NF-κBsignallingpathwaysaccompaniedbytheactivationofMMP-7, therebysuggestingthefeasibilityoftargetingHMGB1forthetreatmentoftumour metastasis.PartIIRedoxStatusofHMGB1PerformsaDualRolein AngiogenesisofColorectal CarcinomaAim:Theobjectiveofthisstudyistoexplorewhetherthedifferentredoxstatesof HMGB1(high-mobilitygroupbox1)haveseparateeffectsontumorangiogenesis.Methods:TomeasuretheexpressionofVEGF-Aandangiogenicpropertiesoftheendothelialcells(ECs),at-HMGB1ords-HMGB1wasaddedtocellmedium,further withtheirspecialinhibitors(DPH1.1mAband2G7mAb)andantibodiesof correspondingreceptors(RAGEAbandTLR4Ab).Also,aco-culturesystemwas appliedtomimictumormic