人肺腺癌紫杉醇耐药细胞株的建立及其生物学特性研究

210002,,,,,,:SPC2A1/Taxol,:,SPC2A1(SPC2A1/Taxol)MTT;;;(mdr1)P2(P2gp):MTT,SPC2A1/Taxol(IC50)SPC2A1759146,TaxotereNVBADM,VP216HCPT,DDPGEM1132;;G0+G1,SSPC2A1P2gp,SPC2A1/TaxolP2gp:SPC2A1/Taxol,,P2gp;;;:R73412:A:1009-0460(2006)09-0657-05Establishmentandcharacteristicsofapaclitaxelresistanthumanlungadenocarcinomacellsubline(SPC2A1/Taxol)YANGAi2zhen,QIANJun,QINShu2kui,CHENHui2ying,LEMei2zhao,XUHai2jun,LIUWen2hu.PLACancerCenter,the81stHospital,Nanjing210002,ChinaAbstractObjective:Toestablishanewpaclitaxelresistanthumanlungadenocarcinomacellsubline(SPC2A1/Taxol)andinvestigateitscharacteristics.Methods:Apaclitaxelresistanthumanlungadenocarcinomacellsubline(SPC2A1/Taxol)wasdevelopedbyintermittentexposuretograduallyincreasingconcentrationofTaxolfromtheparentcelllineSPC2A1invitro.Themultidrugresist2anceofSPC2A1/TaxoltoanticanceragentswasevaluatedbyMTTassay.Themorphologicalfeaturesandthecelluarultrastructurechar2acteristicswereobservedrespectivelybylightmicroscopyandtransmissionelectronmicroscopy.Thedistributionofitscellcyclewasde2tectedbyflowcytometry.TheexpressionofP2glycoprotein(P2gp)wasdetectedbyimmunocytochemistrystaining.Results:SPC2A1/Taxolcellswereof7591462foldresistancetoTaxolanddisplayedhighresistancetoTaxotere,NVB,ADM,butlowresistancewithVP216andHCPT.Nocross2resistancewasobservedtoDDP,GEMandElemeneemusion.Comparingwiththeparentcells,themulti2plicationtimeofSPC2A1/Taxolcellswasprolonged,theirmorphologywasnotchangedobviously,andtheproportionofcellsin(G0+G1)2phasedecreasedwhilethoseinS2phaseincreased.TheexpressionofP2gpinresistantcellswassignificantlyhigherthanintheirparentcells.Conclusions:SPC2A1/Taxolcellsublineisatypicalmultidrug2resistantcelllinewhichhasbasiccharacteristicsofdrug2resistancecells.ItwassupposedthatthemultidrugresistanceofSPC2A1/TaxolwasrelatedtotheoverexpressionofP2gp.KeyWordsHumanlungadenocarcinomacell;Multidrugresistance;Paclitaxel;Cellline,,,2/3,(MDR)2090,,,,[1],,,,75620069119ChineseClinicalOncology,Sep.2006,Vol.11,No.9©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(Paclitaxel,Taxol,),(Taxotere,),(NVB,),(VP216,),(DDP,),(ADM,),(GEM,),(HCPT,)(Elemeneemusion,)(MTT,Fluka),P(JSB21)SP11211211SPC2A1/TaxolSPC2A1Taxol24,0101g/ml,40196g/ml,,,,,,20,1128g/mlTax2ol,SPC2A1/Taxol11212SPC2A1SPC2A1/TaxolSPC2A1SPC2A1/Taxol,310104/ml,1ml24,375%CO2,3,7,Patterson[2]:Td=Tlg2/lg(N/N0),Td,N0,N,T11213MTTSPC2A1SPC2A1/Taxol510104/ml100l/40,375%CO2,24hTaxol510(10%RP2MI21640),4,100l,SPC2A1SPC2A1/Taxol1,96h,510mg/mlMTT20l,4h,,100l0104mol/L,,570nm,,,,(IC50)3=(1-OD/OD)100%;RI=IC50/IC5011214SPC2A1SPC2A1/Taxol,11215SPC2A1SPC2A1/Taxol0102%EDTA,2,4%1%,11216(FCM)SPC2A1SPC2A1/Taxol,,PBS2,70%4,,PBS2110108/L,PI20min,(200),(FACSCalibur)11217P2gpSP,,P2gp,4,,DAB:P2gp,25%,25%50%(+),50%75%(++),75%(+++)11218€xs,SAS811t2211SPC2A1SPC2A1/Taxol331502111441114110,1132(P=01016),212SPC2A1/TaxolMTTSPC2A1Taxol85620069119ChineseClinicalOncology,Sep.2006,Vol.11,No.9©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(RI)759146(P010001)SPC2A1/Taxol36,SPC2A1/Taxol,1,SPC2A1/TaxolNVBTaxotereADM,VP216HCPT,DDPGEM1SPC2A1SPC2A1/Taxol50%IC50(g/ml)SPC2A1SPC2A1/Taxol(RI)Taxol01003701001121810136759146Taxotere0100047010002701400108851106NVB01000650100030514411348369123VP21621931120712311862147DDP01470122015101241109ADM0100520100370122010542131GEM01190106011801030195HCPT21170140410001961184Elemene2emusion2815661132719041860198213SPC2A1,,,;,,,(1),SPC2A1/TaxolSPC2A1,,2214SPC2A1SPC2A1/TaxolSPC2A1,SPC2A1/Taxol,SPC2A1/TaxolG0+G1,S1SPC2A1(A)SPC2A1/Taxol(B)(200),G2+M(2)2SPC2A1SPC2A1/Taxol(%)G0+G1SG2+MSPC2A1691221916811110SPC2A1/Taxol541263014615127215P2gp,SPC2A1P2gp,,SPC2A1/TaxolP2gp75%,33(MDR),,[3],MDR,P2gpMDR,P2gp170kD,,ATPP2gp,[4]20SPC2A1/TaxolSPC2A1/Taxol759146,95620069119ChineseClinicalOncology,Sep.2006,Vol.11,No.9©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(A)SPC2A1/Taxol(B)(7000)3SPC2A1(A)SPC2A1/Taxol(B)P2gp(200),,,S,;SPC2A1/Taxol36,,SPC2A1/Taxol,NVB,8369123,,,ADM,VP216HCPT,DDPGEMSPC2A1P2gp,SPC2A1/TaxolP,[526],24SPC2A1SPC2A1/TAX1216;ADM(VDS)VP216,52FUDDP;P2gp,SKOV3/Taxol2PSK2OV3/Taxol225[7],,[8]LoVo/Adr,,MDR1mRNA,MDR1,,,SPC2A1/Taxol06620069119ChineseClinicalOncology,Sep.2006,Vol.11,No.9©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[1]MozzettiS,ferliniC,ConcolinoP,etal.Classbetatubulinoverexpressionisaprominentmechanismofpaclitaxelresistanceinovariancancerpatients[J].ClinCancerRes,2005,11(1):298-305.[2],,,.27721/Adm[J].,2000,19(8):748-751.[3]UedaK,YoshidaA,AmachiT.Recentprogressinp2glycopro2teinresearch[J].AnticancerDrugRes,2002,14(2):115-121.[4]ThomasH,ColeyHM.Overcomingmultidrugresistanceincanc2er:anupdateontheclinicalstrategyofinhibitingp2glycoprotien[J].CancerControl,2003,10(2):159-165.[5],,,.[J].,2003,43(2):121-122.[6],,,.SPC2A1/TAX[J].,2004,44(5):23-