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:2018-11-20

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1、AbstractMalignanttumorisoneofthemaindiseasesthatthreatenhumanhealth.Atpresent,themaintreatmentmethodsincludeoperation,chemotherapyandradiotherapybutwithsideeffectsandpoorefficacy.Therefore,searchforsideeffectsofsmall,safeandefficienttreatmentisanimportanttrendofantitumorresearch.Ursolicacid(UA)

2、isoneofthenaturalpentacyclictriterpenecompoundsandhasmanybiologicalfunctions.RecentstudieshavefoundthatUAhasantitumorandenhancetheimmunefunction,however,themechanismsofUAonanti-tumorstillnotclear.ThispaperstudiestheUAonproliferationinhibitionandinductionofapoptosismechanismoftumorcellinvitro,MA

3、PKsignalpathwaysandthechangesofcytokineroleinapoptosisinducedbyUA,UAtumorimmuneregulationinvivoandasimmuneadjuvantenhancedimmunogenicityoftumorcellvaccine.ThisstudyprovidesreliableexperimentalevidenceforfurtherapplicationdevelopmentofUAinthefieldofantitumor.Themaincontentsandconclusionsofareasf

4、ollows:1.AntiproliferativeandinducesapoptosismechanismsofUAincervicalcarcinomacelllinesToinvestigatetheeffectofUAoncellproliferationandapoptosisinduction.MTTassaywasusedtomeasuretheproliferationofHeLacells;flowcytometry(FACS)wasusedtoanalysisthechangesincellcycleandapoptosisofHeLacells;Westernb

5、lotwasdetectedCaspase-3,Caspase-8,Caspase-9,CytochromeC,Bcl-2,Bcl-xL,Bak,Baxproteinexpression;Real-timePCRdetectedtheexpressionofHPVE6andE7geneoncervicalcancerHeLaandSiHacells.TheresultsshowedthatUAinhibitedtheproliferationofHeLacellsinadosedependentmanner;theproliferationratewas91.80±3.26%afte

6、r48hof40μMUA,IC50value was9.54±3.51μM.MeasuredresultsshowthatUAarrestHeLacellcycleinG0/G1phaseby flowcytometry,theresultmostsignificantof40μMursoliceffect48h.UAinducedapoptosisofHeLacellsinadosedependentmanner,theearlyapoptosisrateswere74.30±5.26%and83.87±2.27%after40μMand60μMUAafter48hrespecti

7、velyandsignificantlyhigherthanthatincontrolgroupwas2.20±1.15%(P<0.01).WesternblotresultsshowedthatCaspase-3andCaspase-9activationofHeLacellsinducedbyUAandcleavedformsofCaspase-3andCaspase-9wereobserved;theexpressionofCytochromeCpr

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