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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
