About rhuCD40L on human colon cancer transplanted into nude mice the treatment of毕业论文翻译.docx
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About rhuCD40L on human colon cancer transplanted into nude mice the treatment of毕业论文翻译.docx
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AboutrhuCD40Lonhumancoloncancertransplantedintonudemicethetreatmentof毕业论文翻译
AboutrhuCD40Lonhumancoloncancertransplantedintonudemicethetreatmentof
Author:
ZHUXiao-KunMuXiangming,whenNorthKoreaZhouGuangjun
[Abstract]Objective:
TostudyinvivoexperimentsconfirmedrhuCD40Lthetreatmentofhumancoloncancer,whichfurtherexploretheanti-tumormechanism.
Methods:
subcutaneousinoculationofhumancoloncancerHT-29humancoloncarcinomacellmodelestablishedafterthe20nudemicewererandomlydividedintotwogroups,thetreatmentgrouptorhuCD40Lforsubcutaneousinjection,subcutaneousinjectionofnormalsalineasthecontrolgroup,regularlyobservedinnudemicelivingconditionsandmeasuredtheirbodyweight,miceweresacrificed5weeksaftertreatmentandtakethetumor,measuringtumorsizeandtumorweightgroups,andcomparetheresults.
Results:
All20tumorformationinnudemice,allmiceintheexperimentwereingoodlivingconditions,noonediedinnudemice,thecontrolgroupandtreatmentgroupbetweentumorsizeandtumorweightdifferences,thedifferencewasstatisticallysignificant(P<0.05.
Conclusion:
rhuCD40Lonhumancoloncancerhaveatherapeuticeffect.
[Keywords]CD40,CD40ligand,synthetictrimericCD40L,nudemice,coloncancer
CD40LontheexpressionofCD40intumorcellswithawiderangeofgrowthregulation.Alargenumberofstudieshaveshownthat,CD40LandCD40expressioninthetumorcandirectlyinhibittheinteractionofavarietyoftumorgrowth.Coloncancerisacommonmalignancyinthegastrointestinaltract,inChinainrecent20years,especiallyinlargecities,morbidityandmortalityhaveincreasedyearafteryear.thetreatmentofcoloncancerisstillsurgicalresectionisthepreferredtreatment,butbecauseoflocalrecurrenceandmetastasisratewashigherafter5-yearsurvivalrateisstillislow.Thecurrentneedtofindabettermeansofadjuvanttherapytoimprovesurvivalinpatientswithcoloncancer,sothisexperiment,nudemiceasacarrier,usedhumancoloncancercelllineHT-29humancoloncancertransplantedintonudemicetoestablishthemodeltoResearchrhuCD40Lanti-tumoreffect.
1Materialsandmethods1.1cells:
HT-29humancoloncancercelllines,providedbytheShanghaiCellBankofChineseAcademyofSciences.1.2Experimentalanimals:
SPFlevel20nudemice,4to5weeksold,weighing16to20grams,arefemale,bytheChineseAcademyofSciencesShanghaiExperimentalAnimalCenterofSoochowUniversitywerehousedinSPFLaboratoryAnimalCenter.
1.3Reagentsanddrugs:
McCoy’5AwerepurchasedfromGiBcocompany;rhuCD40LpurchasedfromCaymanChemicalCompany.1.4Cellcultureandpassage:
thefollowingoperationsareperformedinsterilecellculturechamberforcleanworkbench.HumancoloncancerHT-29cellsinMcCoy’5Amedium(containing10%fetalbovineserummaintainedat37℃,5%CO2incubatorboxculture,adherentmonolayercellgrowth,adherentcellfusiontobe70%to80%ofthetrypsin(containing0.02%EDTAdigestionandpassage.
1.5cellswerecollectedandinoculatedinnudemice:
thecellsinthelogarithmicphaseafterdigestionwithtrypsinMcCoy’5Awithserum-freeculturemediumdilutedcellsuspension,centrifuged,thesupernatantwasdiscarded,andthentoserum-freeMcCoy’5Atrainingrepeatedwashing,centrifugationthreetimes,supernatantculturemediumwithserum-freeMcCoy’5Adilutedcellsandadjustcellconcentrationto3*107/ml,totakethecellsuspension0.1ml/injectioninnudemiceonlyweretherootsoftherightupperlimbbackskin.
1.6grouptherapy:
tobevisibletumorgrowninnudemiceaftertheinoculationsite,the20nudemicewererandomlydividedintotwogroupsof10each.GroupA(controlgroup:
saline0.5ul/g/dayasasubcutaneousinjectionaroundthetumor,1time/day,3daysfortreatment,weeklytreatment,atotaloffiveweeks,Bgroup(treatmentgroup:
therhuCD40L0.5ug/g/dayasasubcutaneousinjectionaroundthetumor,1time/day,3daysfortreatment,weeklytreatment,continuoustreatmentfor5weeks.
1.7,respectively,aftertreatment1,3,7,14,28,35eachdaywithaverniercalipermeasurementoftumorsize,andrepeatthemeasurementthreetimesandaveragetumorvolumeinthefollowingformula:
V=ab2π/6a:
tumornodulemaximumdiameterb:
thecrossbythetumornodules,thelastmeasurementaftertheremovalofnudemiceweresacrificed,removalofthetumorweightduringtreatmentbutalsoacontinuousobservationofnudemicediet,urine,mental,activityandweightlossandsoon.
1.8StatisticalMethods:
Alldatais+-ssaid,usingSPSS15.1statisticalsoftwarepackageforstatisticalanalysistotestthelevelof0.05.2results2.1experiment20micewereinHT-29humancoloncancercellswereinoculatedsubcutaneouslywith4~5dafterinoculationsiteoccurinabout0.4cmindiameterthesizeofthetumornodules,tumorformationwas100%duringtreatmentwereallnudenoobviousactivity,lossofappetite,nausea,vomiting,reflectingtheslow,listlessness,weightlossandsoon.endoftheexperimentdiedofnaturalcauseswhennoonenudeexperimentnudepicturesinFigure1.
2.2duringthetreatmentoftumorgrowthtrendshowninFigure1(basedonthecourseoftreatmentthetumorvolumemeasureddata
2.3Aftertreatment,tumorvolumemeasuredvalues?
?
inTable1.
Groupsofanimals(ntumorvolume(cm3Agroup102.028±0.047BGroup100.767+-0.007t=77.18,p<0.05
2.4Aftertreatment,tumorweightineachgroupisshownintable2.
Groupsofanimals(ntumorweight(gAgroup101.992+-0.224Bgroup100.982+-0.410t=68.412,p<0.05
3DiscussionCD40isatumornecrosisfactorreceptor(tumornecrosisfactorreceptor,TNF.R)familymemberexpressedinBlymphocytes,monocytes,hematopoieticstemcells,dendriticcells(DCs),endothelialcells,epithelialcellsandothersurfaces.CD40ligand(CD40ligand,CD40L),alsoknownasCD154,isatumornecrosisfactor(TNF)familymembers,mainlybyactivatedCD4+Tlymphocyteexpression.CD40LinteractionwithCD40caninhibitbreastcancer,ovariancancer,cervicalcancer,bladdercancer,non-smallcelllungcancerandsquamouscellcarcinomagrowth,theroleandCD40Lcaninducecellcyclearrestoftumorcellsand/orapoptosis.CD40LDCscaninducecellphenotypeandfunctionofmaturity,youcanincreasetheexpressionofcostimulatoryfactorsandcytokinestoenhancetheimmunogenicityofepithelialtumorsandmanymodelshavedemonstratedCD40Lstimulateanti-tumorimmuneresponsecapabilities.Rongandotherreports,usingthesametimeencodingcarcinoembryonicantigen(CEA)andCD40LtrimervaccinesuccessfulinCEAtransgenicmiceinducedratcoloncancerandeffectiveprotectiveimmuneresponse,intheprocess,observedwithoutantigenstimulationofTcellsandactivationofDCs[3]inthehomologousratsolidtumormodel,injectiondirectlyintotheADV-CD40LmicewereinoculatedsubcutaneouslyintotheB-16melanoma,MCA-1sarcomaorCT-26coloncancer,areinmostofthetumor-bearingmicereceivedtheeffectoftumorregression[4]usingencodingCD40Lreplication-defectiveadenovirus(ADV-CD40L),canmakechroniclymphocyticleukemiaBcellsandfollicularlymphomacellstoenhancetheoriginalco-stimulatorymoleculeexpression[5][6]andthesecellstheabilitytostimulateTcellproliferationweresignificantlyenhanced[7].
rhuCD40LissolublerecombinanthumanCD40receptorligand,toincreasethestabilityoftheisoleucinezipperintheform[8].vitroexperimentsonbreastcancercelllinesconfirmedrhuCD40Lsignificantlyinhibitedthegrowthoftherole,butalsoinvivoexperimentsalsoconfirmedasignificantincreasethesurvivalrateofSCIDmice[9].Studieshaveshown,rhuCD40LcaninhibithumanCD40+ovariancancerSCIDmousexenografttumorgrowth,rhuCD40LalsoincreasetheamountofCDDPthantheeffectoftreatment[10].
Studyathomeandabroadbasedonthefield,studythenudeasacarrier,confirmedbyanimalexperiments,rhuCD40Lcaninhibitthegrowthofhumancoloncancer,anditsanti-tumormechanismneedsfurtherstudy.First,thelackofnudemicewithcongenitalimmunityoftheexperimentaldruginthetreatmentofimmunefunctionlessinterference.Secondly,theexperimentusingtumoraroundthesubcutaneousrouteofadministration,thewayaroundthroughtheabsorptionofdruginthetumordirectlyonreceptorswithinthetumor,shortertreatmentTimetoplayastrongerroleinthedruginthedrugtreatmentafterbymeasuringtumorsizeandtumorweight,tumorconfirmedthetherapeuticeffectofdrugsonthisexperimentalsoconfirmed,rhuCD40Ldietofexperimentalanimals,spirit,activity,bodyweight,urine,etc.isless,soyoucanenvision,rhuCD40Lforhumananti-tumortreatment,thebodymaynotresultinsignificantdamagetotherole,so,rhuCD40Lasanadjuvanttherapyforcancerdrugsinhumantreatmentofcoloncancerimmuneopenupbroadprospects.
References[1]PaulieS,Rose‘nA,Ehlin-HenrikissonB,etal.ThehumanBlymphocyteandcarcinomaantigen,CDw40,isaphospho-proteininvolvedingrowthsignaltransduction.JImmunol.1989;142:
590-595.
[2]ArmitageRJ,SatoTA,MacduffBM,etal.IdentificationofasourceofbiologicallyactiveCD40ligand.EurJImmunol.1992;22:
2071-2076.
[3]XiangR,PrimusFJ,RuehlmannJM,eta1.Adual-functionDNAvaccineencodingcarcinecmbryonicantigenandCD40ligandtrimerinducesTcell-media-tedprotectiveimmunityagainstcoloncancerincarcinoembryonicantigen-transgenicmice[J].JImmunol,2001,167(8):
4560-4565.
[4]TongAW,StoneMJ.ProspectsforCD40-directedexperimentaltherapyofhumancancer[J].CancerGeneTher,2003
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