简介：ThepresentstudyisaimedatstudyingthegeneforTIMP-3,amammaliantissueinhibitor,byconstructingarecombinanteukaryoticcellvectorforgenetherapyinhumanbreastcancer.WeobtainedtheTIMP-3genefromthehumanplacentbyRT-PCR.TIMP-3genewassubclonedintopcDNA3.1vetorfrompMD18TvectorbymeansofgenecloningtoconstructpcDNA3.1recombinantvector.HumanbreastcancercelllineMDA-MB-453wastransfectedwithpcDNA3.1-TIMP3recombinantvectorusinglipofectaminereagent.ThentheexpressionofTIMP-3andtheeffectonthemetastasisofMDA-MB-453wereexamined.ThecorrectconstructionofpcDNA-TIMP3wasidentifiedbymeansofrestrictionenzymeanalysis,PCRamplicationandnucleotidesequencing.WesternblottingshowedthatthetransfectedcellswereabletoexpressTIMP-3,indicatingthatourconstructionofthepcDNA-TIMP3eukaryoticexpressionvectorwasconstructedsuccessfully.OurexperimentsfurtherindicatedthatthepotentialofmetastasiswassignificantlyreducedforthetransfectedcelllineMDA-MB-453.