人乳腺癌細胞系經(jīng)誘導分化后差異表達基因的初步研究.pdf

1、南開大學碩士學位論文人乳腺癌細胞系經(jīng)誘導分化后差異表達基因的初步研究姓名：邢德印申請學位級別：碩士專業(yè)：七年制臨床醫(yī)學指導教師：馮玉梅毛慧生19990501南日K_.大學石泣二卜學1立論文PreliminaryStudyofDifferentialE沖ressedGeneinMCF7CellLineTreatedbyalltransRetinoicAcidGraduate:XingDeyinDepartment:ClinicalMedi

2、cineSupervisor:FengYumeiMaoHuishengLiuHongyinAbstractObjective:DevelopmRNAdifferentialdisplaypolymerasechainreaction(DDPCR)methodwithsilverstainingStudytheeffectofinduceddifferentiationinbreastcancercelllineMCF7treatedby

3、alltransretinoicacidandisolatedifferentiationrelativegenes.Methods:TotalRNAareextractedfromMCF7celllinetreatedbyalltransretinoicacidandcontrolones.AfterreversetranscriptionreactioncDNAsareamplifiedbyDDPCRusingthreeanchor

4、edprimersandfourarbitraryprimers.TheDNAbandsonpolyacrylamidegelsaredisplayedbysilverstainmethodthenthedifferentialdisplayedDNAbandsarevisuallyrecoveredfromgelsandreamplifiedbyPCR.Reamplifiedproductsareoftenamixtureofsimi

5、larsizewhicharedeterminedbysinglestrandconformationpolymorphisms(SSCP)gels.Results:ThereareobviousdifferencesincharacterizationofcellbiologyandgeneexpressionbetweenhumanbreastcancercelllinesMCF7beforeandafteralltransreti

6、noicacidtreatment.SeveralcDNAfragmentsexpressedaftertreatmentareisolatedoneofwhichis313bpsizeandhasbeendeterminedbySSCPgels.Conclusion:l.Thedevelopednonradioactivedifferentialdisplaytechniquewithsilverstainingisasimplese

7、nsitiveandefficientmethodforisolatingdifferentialexpressedgene2.AI]transretinoicacidplaysanimportantroleinregulatingthedifferentiationrelatedgeneexpression.Investigatingthisprocesswillhelpustoexplainthemechanismofinduced