熊果酸抗腫瘤及作為小鼠肝癌細(xì)胞疫苗佐劑的作用研究

熊果酸抗腫瘤及作為小鼠肝癌細(xì)胞疫苗佐劑的作用研究

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時(shí)間:2018-11-19

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1、AbstractMalignanttumorisoneofthemaindiseasesthatthreatenhumanhealth.Atpresent,themaintreatmentmethodsincludeoperation,chemotherapyandradiotherapybutwithsideeffectsandpoorefficacy.Therefore,searchforsideeffectsofsmall,safeandefficienttreatmentisanimportanttrend

2、ofantitumorresearch.Ursolicacid(UA)isoneofthenaturalpentacyclictriterpenecompoundsandhasmanybiologicalfunctions.RecentstudieshavefoundthatUAhasantitumorandenhancetheimmunefunction,however,themechanismsofUAonanti-tumorstillnotclear.ThispaperstudiestheUAonprolif

3、erationinhibitionandinductionofapoptosismechanismoftumorcellinvitro,MAPKsignalpathwaysandthechangesofcytokineroleinapoptosisinducedbyUA,UAtumorimmuneregulationinvivoandasimmuneadjuvantenhancedimmunogenicityoftumorcellvaccine.Thisstudyprovidesreliableexperiment

4、alevidenceforfurtherapplicationdevelopmentofUAinthefieldofantitumor.Themaincontentsandconclusionsofareasfollows:1.AntiproliferativeandinducesapoptosismechanismsofUAincervicalcarcinomacelllinesToinvestigatetheeffectofUAoncellproliferationandapoptosisinduction.M

5、TTassaywasusedtomeasuretheproliferationofHeLacells;flowcytometry(FACS)wasusedtoanalysisthechangesincellcycleandapoptosisofHeLacells;WesternblotwasdetectedCaspase-3,Caspase-8,Caspase-9,CytochromeC,Bcl-2,Bcl-xL,Bak,Baxproteinexpression;Real-timePCRdetectedtheexp

6、ressionofHPVE6andE7geneoncervicalcancerHeLaandSiHacells.TheresultsshowedthatUAinhibitedtheproliferationofHeLacellsinadosedependentmanner;theproliferationratewas91.80±3.26%after48hof40μMUA,IC50value was9.54±3.51μM.MeasuredresultsshowthatUAarrestHeLacellcycleinG

7、0/G1phaseby flowcytometry,theresultmostsignificantof40μMursoliceffect48h.UAinducedapoptosisofHeLacellsinadosedependentmanner,theearlyapoptosisrateswere74.30±5.26%and83.87±2.27%after40μMand60μMUAafter48hrespectivelyandsignificantlyhigherthanthatincontrolgroupwa

8、s2.20±1.15%(P<0.01).WesternblotresultsshowedthatCaspase-3andCaspase-9activationofHeLacellsinducedbyUAandcleavedformsofCaspase-3andCaspase-9wereobserved;theexpressionofCytochromeCpr

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