TPA对人食管癌Eca109细胞增殖和细胞核形态的影响.docx

TPA对人食管癌Ecal09细胞增殖和细胞核形态的影响TPA对人食管癌Eca109细胞增殖和细胞核形态的影响TPA对人食管癌Eca109细胞增殖和细胞核形态的影响作者：郭玉忠，贾琳静，部娜，田鑫，谢敏，张莉蓉，乔海灵【摘要】目的研究佛波(12-0-tetradecanoy1phorbo113-acetate,TPA)对人食管癌细胞的影响。方法细胞增殖试验采纳MTT法和细胞计数的方法，细胞核形态的测量采纳计算机协助的图像分析系统，分别测量计算细胞核面积(NA)、核周长(NP)、最大直径(DmaX)、最小直径(Dmin)、核浆比(NPA)溟、核轴比(NAR)、核形态因子(NR掰F)等。结果TPA没有抑制人食管癌Ecal09细胞增殖和变更细胞核形态漱。结论TPA对人食管癌Eca109鹰细胞可能没有促分化作用。【关键词】佛波酯；食管癌细胞；细胞核形T态EffectsofTPAo跋nproliferationandnUClearmOrPhOmeIryo贽fhumanesophagealca撕ncerEcalO9cells【A殛bstractObjective湾SToinvestigatethe溶effectsof12-0-tetr揣adecanoy1phorbo1-13-acetate(TPt)onhu疙manesophagealcance垩rEcalO9celIs.MethodsThesensitivityoVlfTEAtoEca109ce11sw酥asdeterminedwithMTTassayandcountingc侔el!numbers.Afterth稼ecelIswerestaincdw否ithHEIhCmCaSUremCntsofnuc1earmorpho破metrywereperfor11ed咫WilhComPUIer-assisIedimageanalysissy籀stem.ResultsTPdi辨Clnotinhibitthecel1咬sproliferationandd枭CcreasethenucIears哪ize.ConclusionsTP麽sophageaIcanccriso绐neofthemostco11unons筠olIdtumorsinChina.AmaynotinduceEcalO铢9celIstodifferenti遍ate.Keywords12钻-O-tetradecanoyIph海orbol-13-acetate(T嫂PA)：esophagealcanc每ercellline；nuclearmorphometryHumaneDuetoitshighmetast松aticpotentialandthefrequentonsetofre翩Sistancetochemothe噬rapy,itisoncofthcmajorcausesofdeathb衡yneopIasiainthecou谈ntry.Thepresentche轿Sesagentsthatareus剑cells.OntheotherhaOndUlardifferentiatio站motherapyofcanceru辛ua11ytoxiCtonorma1羚inductionofcel1孝nmaysupplementtheu理seofnon-cytotoxicdkrugsinsevera1formsOfneoplasia,1iketh绅Csuccessfuluseofal1-transretinoicaci运d(?TRA)inthetreatm镂entofacutepromyelo番Cyticleukemia.Pla-niditerpinephorbol笛esters,particular1荀ythephorbolesterl2幺-O-tetradecanoylph灵orbol-13-acetate(T哈tpotenttumorpromotfeetsrangefromstimSthroughactivating源Oducingalterationsesandconstituents.臼tetHedifferentiati调Ilsincultureandinv睐PAcaninducediffere殃PA)»areamo11gthemos鸣erSknownandthEiref祸UlatingDN-Asynthesi蝮geneexpressiontopr栽WivinmembrancpropcrtiTPcanalsopotentiaonofvarioustumorce禁iVoofhumanbEIngs.Tntiationandinhibit彝thegrowthofanumber臼ofmalignantcelltyp宋es,includingmyeloi调dleukemia1,breast2»prostate3,c吉olon4,andbrain5英.OtherreportshoweHthatTPAcouldnotin航rcinomacelIline花6.BccauseofitseffeareinterestedinitYfPhagealcancercells抖ffectsoP.onpro1i孩morphometryofhumanducehumanhepatoca管todifferentiation听iciencyi11vivo7,”,诸Seffectsonhumaneso7.Here»Wereportthee莫esophagealcancerEca!09cel1s.1Mater潭ialsandmethodsChedfromSigmaChemica抠emicalsTPApurchas碳ICo.Wasdissolvedin哧ferationandnuc1ear溥ethanolandstoredat蹇-204C.Thestocksolut炷ionswerediIutedwit鳗hHanksbuffercdsaltsolution.Thefinal邙Concentrationofeth虢anolintheincubatio15.nmixtureswas%orles皎s.CellGrowthStud狡iesHumanesophagea½Icancercel1IineEca109WaSkindiyPrOVid侵edbyprofessorBaiji屉ngxiu,ZhengzhouUniversity.Thecellswe巳remaintainedinRPMIr1640mcdiumwithl%fetalbovineserum(FB烟S)Inthepresenceofa娱ntibiotics,penici1摄lin(100unitsml)an3dstreptomycin(100u旖nits/ml).Allcellsw醛ereculturedat37*Cinahumidifiedatmosph咖erecontaining5%C02鳗.Themediuinwasrepla好Cedafter3days.Thec呈e11Swereroutine1ysUbculturedtwiceaweek.Cellswercplate秘(Iineitherflasks(25讫cm2)or96-weHplateX"s.TPAwasadded24hou迂rsafterseeding»who筏nthecelIswereattac埠hedtothesurfaceoft廉hecontainers.AnequivalentvolUmeofethanolwasaddedtothec饬ontro1SandhadnoeffXectongrowth.Afterr萱emovingIhecellsfroIntheflasksbytrypsiXInizatiOntheywereco退Untedusingahemocyt杼Ometer,fixcdwith70黑%ethanolandstained囱byWhenthecelIswere榭platedintoa96-wel1缗microplatethecult灭uremediumcontainin硅gdifferentconcentr柱alionSofTPAWaSrCPlaced24hourslater.After3-5daysofincub苴ation,Cellviabilit溅ywasevalUatedbyMTT诫assay.Nuclearmor漆PhometrymeasurementsMeasurenientsof忍nucIearmorphometry恻inc1udingnuc1eararea(NA)»nucIearperimeter(NP),maximaid响iameter(Dmax),minimaldiameter(Dmin),得nucIearplasmaratio哩(NPR)»nuclearaxisr惺atio(NAR),nucIearrOundnessfactor(NRF的)»wereperformcdwit水hcomputer-assisted磋imageanaIysIssystemMPIS-500(QingPin愍gImageEngineeringC葩ompany,TongjiUniversity).Twofieldswe恪reselectedatrandom眈fromcachcaseforthc探measurements.Eachf睛ie1dwasviewedunder齿a100oi1-immension亮1cnsanddisp1ayedon魔theserecnofacoIorvyideoimageprocessor屐.TheimagesofSOnuc1首eiofthetumourcelIs宗Perfieldwcreoutlin隶edwithadrawingpen9andthedatawereproc禹Cssedbyacomputerin悯thesystem»whichcal铉Culatedtheparamete激rs.Intheendallthem蛾Canvaluesofeachcasewcrccalculated.StatisticalAnalysi网SThedatawereexpre萧SsedasmeansSD.Sta骆tistJcalanalysiswa骼SPerfonnCdbySPSSSoftware.Thecriterio1nforstatisticalsig约nifiCancewastakena倾sP2ResultsEffect-j-OfTPAoncel!Growth.ThegrowthofEcal09c雷Cllswasnotarrested骄byTPAatnoneytotoxi峋Cconcentrations.As押ShowninTableland,TPAevenincreasedtheEcancercel!numbers»(IDthoughtherewerenosignifiCanc