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1a21b1b31．a．b2640052．2640033．210095A549、SGC－7901、MCF－7PANC－14MTT、sephadexG－50DEAE－52Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE33。343MCF－7A5490．02、0．04、0．06、0．08、0．10mg/mLⅡ－ⅠDⅡ－ⅠEA549Ⅱ－ⅠCⅡ－ⅠDMCF－7、SGCPANC－1Ⅱ－ⅠEⅡ－ⅠC393%β－31．52%～32．67%。3。Q956APurificationandantitumoractivityofpolysaccharidesfromThelenataanaxLIUHongmei1aZHOUXiaoqiu2YAOYanan1bJIANGAili1bWANGChanghai31．a．SchoolofOceanb．CollegeofLifeSciencesYantaiUniversityYanta264005China2．YantaiInstituteofCoastalZoneＲesearchChineseAcademyofSciencesYantai264003China3．CollegeofＲesourcesandEnvironmentalSciencesNanjingAgriculturalUniversityNanjing210095ChinaAbstractInvitroantitumoractivitiesofpolysaccharidesfromThelenataanaxwereinvestigatedonfourcancercellsincludingA549SGC－7901CF－7andPANC－1usingMTTmethod．ContinuoushydrolysisbytrypsinandpepsinfollowingpurificationthroughsephadexG－50andDEAE－52respectivelythreerefinedpolysaccharidesnamedⅡ－ⅠCⅡ－ⅠD、Ⅱ－ⅠEwereobtained．Theresultsshowthatthethreerefinedpolysaccharideshavesignificantlystrongerantitumoractivitiesthancrudepolysaccharideswithpositiverela-tiontotheirmassconcentrationsandthattheyhavethestrongestinhibitioneffecttoMCF07andweakerantitumoractivitytoA549．Atexperimentalconcentrations0．020．040．060．080．1mg/mLⅡ－ⅠDandⅡ－ⅠEhavestrongerinhibitioneffectonA549thanⅡ－ⅠCandⅡ－ⅠDhashigherantitumoractivitiesonMCF－7SGCandPANC－1thanⅡ－ⅠEandⅡ－ⅠC．Analysesonthethreepolysaccharidesshowthatactivepolysaccharidesarepyranpolysaccharideswithβ－configurationwithoutpeptideandnucleicacidwithover93%puritycontaining31．52%～32．67%—O—SO3—residues．Themonosaccharidecompositionshowsthethreepolysac-charidesallbelongtoseacucumberchondroitinsulfateSC－CHS．KeywordsThelenataanaxpolysaccharidepurificationantitumoractivityseacucumberchondroitinsulfates、、、、、1－2。22015－09－212016－06－06115031070368。1965—。jmszz.cnSC－CHSD－N－、D－L－SC－FUCL－3－5。6。。4℃60%15%、。。。。。2。、、、。11．1。。D－Man、D－+－GalN、D－+－GlcN、D－、D－Gal、D－GlcUAL－FucSigma。A549、MCF－7、SGCPANC－1。MTTDM-SOSigma－AldrichDMEM、ＲPM-ll640GIBCO。1．2JASCOSANYOMCO－15ACCO2Bio－radAglient1100Aglient6890N。1．31．3．11095%45℃4h2pH1．5400U/g37℃3h。4000r/min10minPartⅠPartⅡ。PartⅠpH7100℃40min4000r/min10minpH7．8400U/g37℃3h100℃40min4000r/min10minpH7．034℃4000r/min15minⅠ。SephadexG－501mL/minⅠ－ⅠⅠ－Ⅱ。PartⅡ10pH7．8400U/g37℃3h100℃40min4000r/min10min34℃4000r/min15minⅡ。SephadexG－501mL/minⅡ－Ⅰ、Ⅱ－ⅡⅡ－Ⅲ。Ⅱ－ⅠDEAE－521mL/min6Ⅱ－ⅠA、Ⅱ－ⅠB、Ⅱ－ⅠC、Ⅱ－ⅠD、Ⅱ－ⅠEⅡ－ⅠF。1．3．21．3．2．1A549、SGC－7901、MCF－7PANC－1ＲPM11640DMEM4043037℃、5%CO2SANYOELECTＲICCo．Ltd。。1．3．2．2A549、SGC、MCF－737℃2～3min1000r/min6minＲPMll6405×104mL－196200μL24h3100μL。24h20μLMTT5mg·mL－14h150μL490nm。inhibitionrateIＲ=1－/×100%。SASV8．2。1．3．3HPSEC。TSKPW50005μL1mL/min25℃206nm。IＲ1～2mgKBrIＲ400～4000cm－1。BaCl27。8。DB22530m×0．32mmi．d．0．25μmFID250℃250℃220℃1mL/min。22．1SephadexG－503Ⅰ－Ⅰ、Ⅰ－ⅡⅡ－Ⅰ1。Ⅱ－ⅡⅡ－Ⅲ。14/mg·mL－1/%A549SGC－7901MCF－7PANC－1Ⅰ－Ⅰ0．530．26±1．22＊＊＊20．20±1．26＊＊＊2．20±0．162．46±0．110．420．94±1．24＊＊15．26±1．35＊＊1．63±0．252．12±0．120．311．68±1．25*11．63±1．23*1．26±0．061．95±0．160．24．06±1．21*6．00±1．11*1．15±0．211．46±0．110．15．40±1．22＊＊3．26±1．12*1．06±0．221．23±0．20Ⅰ－Ⅱ0．528．3±1．32＊＊＊2．33±0．051．05±0．052．02±0．020．426．27±1．34＊＊＊2．27±0．021．01±0．041．21±0．020．321．99±1．26*1．99±0．010．99±0．051．52±0．030．216．51±1．12＊＊1．51±0．020．76±0．060．87±0．040．17．36±1．21*1．36±0．020．53±0．030．65±0．03Ⅱ－Ⅰ0．531．56±1．35＊＊74．81±1．28＊＊61．63±2．28＊＊55．70±3．12＊＊0．427．44±1．23＊＊＊69．98±2．42＊＊＊54．86±2．42＊＊＊47．46±2．57＊＊0．319．73±1．24＊＊55．18±1．25＊＊45．77±2．25＊＊32．65±2．68＊＊0．212．55±1．33＊＊43．86±1．23＊＊22．92±1．23＊＊7．11±1．15*0．17．9±1．12＊＊32．98±1．31＊＊12．03±1．31＊＊3．32±0．52**P＜0．05＊＊P＜0．01＊＊＊P＜0．001。504534。Ⅱ－Ⅰ4SGC－7901Ⅰ－ⅠA549SGC－7901MCF－7PANC－1Ⅰ－ⅡA5493。2．23Ⅱ－ⅠⅡ－ⅠA、Ⅱ－ⅠB、Ⅱ－ⅠC、Ⅱ－ⅠD、Ⅱ－ⅠEⅡ－ⅠF。3Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE1。34。3MCF－7A549。3Ⅱ－Ⅰ0．1mg/mL0．5mg/mLⅡ－Ⅰ3Ⅱ－Ⅰ。aA549bSGC－7901cMCF－7dPANC－13±。。*P＜0．05＊＊P＜0．01＊＊＊P＜0．001。13Ⅱ－ⅠDⅡ－ⅠEA549Ⅱ－ⅠCⅡ－ⅠDMCF－7、SGCPANC－1Ⅱ－ⅠEⅡ－ⅠCⅡ－ⅠEⅡ－ⅠCSGCPANC－1Ⅱ－ⅠCMCF－7Ⅱ－ⅠE。Ⅱ－ⅠCSGC－7901IC500．061mg/mLIC500．1mg/mLIC500．095mg/mLⅡ－ⅠD60430SGC－7901IC500．053mg/mLIC500．044mg/mLIC500．069mg/mLⅡⅠESGC－7901IC500．069mg/mLIC500．093mg/mL。202。Sl8053%51%55%MA－73779%～88%LewisT79566%～79%。2．332．3．1Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE485nm540nm。3。HPSECⅡ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE95．15%、93．23%97．01%203．5kDa、215．6kDa193．4kDa。、。2．3．2Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE8－103500～3400cm－1O—H2962～2850cm－1C—H1510～1670cm－1CO1485～1445cm－1C—H1083cm－1C—O800cm－1890cm－1β－840～853cm－1α－1265cm－1—O—SO3——SO—。3β－—O—SO3—。2．3．3。11－12。Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE332．45%、31．52%32．67%13。Ⅱ－ⅠDⅡ－ⅠCⅡ－ⅠE14。2．3．432。3Fuc、GlcUA、GalNGlcNSC－CHS15－16GalMan39。23Fuc∶GalN∶GlcUA∶GlcN∶Gal∶ManⅡ－ⅠC1∶0．68∶0．65∶0．11∶0．12∶0．11Ⅱ－ⅠD1∶0．96∶1．01∶0．32∶0．15∶0．11Ⅱ－ⅠE1∶0．76∶0．86∶0．02∶0．13∶0．053、Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠE393%3β－31．52%～32．67%。3Ⅱ－ⅠC、Ⅱ－ⅠDⅡ－ⅠEA549、SGC－7901、MCF－7PANC－14。3MCF－7A549。50．02、0．04、0．06、0．08、0．1mg/mLⅡ－ⅠDⅡ－ⅠEA549Ⅱ－ⅠCⅡ－ⅠDMCF－7、SGCPANC－1Ⅱ－ⅠEⅡ－ⅠC。。1．J．20122481151－1159．2．TJ．201410870451945－1948．3YUTAKAKBAＲBABAMKYOKOIetal．Isolationandpartialcharacterizationoffucansulfatesfromthebodywallofseacucum-berStichopusjaponicusandtheirabilitytoinhibitosteoclastogenesisJ．CarbohydＲes20043391339－1346．4．3J．2015412227－232．5ZHANGXWANGJFXULetal．Comparativeanti-tumoreffectsoffucoidanfromtwoseacucumberspeciesonspontaneousmetasta-sisoflewislungcarcinomainmouseJ．FoodSci2012337251－255．6．、D．2010．7DODGSONKPＲICEＲ．