α,-岩藻糖转移酶基因转染对卵巢癌细胞生物学行为影响.docx

α,-岩藻糖转移酶基因转染对卵巢癌细胞生物学行为影响.docx

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α,-岩藻糖转移酶基因转染对卵巢癌细胞生物学行为影响.docx

二、Methods1.ExtractionofHFUT—HandpcDNA3.1:extractedplasmidaccordingthedirectionofcellphecttranstectionkit.to2.Transfectionandresistantcellcolonscreening:RMG—1weretrans—fectedbyHFUT—HandpcDNA3.1screenedtheresistantclonewithG418.withcellphecttransfectionkit.And3.Immunocytochemistry:thetrialwascarriedoutaccordingtothedirectionofSABCkit.4.Observationofcellmorphology:observedcellswithinvertmicroscopeandHEdying.5.Proliferationexperiment:drawngrowthcurvebyMTYandcellcounting.6.Softagarculture:cells14dayslater,counted.wereinoculatedindoublelayeragarplate,and7.AnMysisofcellcyclebyflowcytometry:afterPIdying,cellstectedbyflowcytometry.8.Statisticalanalysiswerede—StatisticalanalysiswasperformedbytheSPSSl1.0package.ProbabilityofP<0.05wasconsideredstatisticallysignificant.Resuits1.ExtractionO{HFUT—HandpcDNA3.12.1mmunocytochemistry:thepositiverateofLewisYexpressedinRMG—I—HishigherthaninRMG—IandRMG—I—pcDNA3.1,P<0.05.3.Observationofcellmorphology:RMG—IandRMG—I—pcDNA3.1growthinlamellarshapeandtheborderlineisregular,buttheborderlineofRMG—I—itisirregularandcellssuperposably.areeasytoshedfromcultureflaskandgrowly.4.GrowthCUlWe:RMG—I—Hgrowsfasterthantheothertwosignificant-5.Softagarculture:rateofcloneformation:RMG—I—His35%,RMG—IandRMG—I—pcDNA3.1were13%and12%separately,P<0.05..气.6.Analysisofcellcycle:comparedwithRMG—IandRMG—I—pcD—NA3.1,percentageofRMG—I—HcellsdecreasesinG1phaseandGophaseandincreasesinG2phaseandinMphase.DiscussionsTheglycocalyxlayerofmammaliancellsisknowntochangeinassociationwithcellularadhesion,recognize,canceration,invasionandmetastasis.TherearesufficientevidencesthatLewisantigensaletumor—associatedmoleculesandcorrelatedtohi。ghgradeandpoorprognosistumors.Wetransferred理1.2一FTgeneintoRMG—Icellsandconstructedcellu—laxmodelexpressingd1.2一FrgeneandLewisYhighlyfirstly.TheexpressionofLewisYwasfoundtoincreaseinRMG—I—Hbyimmunocflochemistry.bi—ologicMbehaviorofwhichchanged.Thereasonsofthatmaybe:(1)modificationsofglycosyltransferasemaketumorcellsescapefromimmunity;(2)negativechargein

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