CELLULAR PROCESSING FLUOROPHORAND DIGITAL FLUORESCENCE MICROSCOPY.pdf

《CELLULAR PROCESSING FLUOROPHORAND DIGITAL FLUORESCENCE MICROSCOPY.pdf》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、JBiolInorgChem2000)5:655±665DOI10.1007/s007750000153ORIGINALARTICLEChrisMolenaar´Jan-MaartenTeubenRobertJ.Heetebrij´HansJ.Tanke´JanReedijkNewinsightsinthecellularprocessingofplatinumantitumorcompounds,usingfluorophore-labeledplatinumcomplexesanddigitalfluorescencemicroscopyReceived:5April2000/Ac

2、cepted:21June2000/Publishedonline:29July2000SBIC2000AbstractThecellulardistributionandprocessingvesiclesinintracellularprocessingofcisplatin-derivedpathwaysoftwoplatinumcompounds,modelingthecomplexes.Immunocytochemicalstudies,usingtheantitumordrugcisplatincDDP)inhumanosteosar-DNPderivative,resu

3、ltedinverysimilarimagesascomaU2-OS)cellsisreported.A[Pten)Cl2]entityobtainedwiththeCFDAconstruct.CFDA-bocahasbeencovalentlylinkedtoacarboxyfluoresceinnon-platinum-containingfluoresceinderivative)wasdiacetateCFDA)moietyandtoadinitrophenylusedascontrol:afaintstainingthroughoutthewholeDNP)moiet

4、y.Thetwodifferentconstructswerecellwasobserved.Cisplatin-resistantU2-OS/PtcellsadministeredtolivingcellculturesthatwereanalyzedshowedstainingpatternsverysimilartotheU2-OSusingdigitalfluorescencemicroscopy.Thenon-flu-cellsusingbothplatinumconstructs.Thisstudyillus-orescentCFDAconstructbecomesfluor

5、escentaftertratesthatonlyaverysmallportionoftheplatinumcellularuptakeandsubsequentacetatehydrolysisbycomplexeventuallyremainsboundtoDNA,asafteresterases,andisthereforesuitabletomonitorplati-24hnosignificantfluorescencecouldbeobservedinnuminlivingcells;theDNPconstructcanbevisu-thenucleus.Cisplatin

6、-derivedcomplexeswithfluores-alizedbyimmunocytochemistryandconsequentlycenttagsaffordanewinsightintothecellularprocess-servesasacontrol.Bothcomplexeswerereadilyinter-ingofthesecomplexesandthereforemaycontributenalizedbythecells,andlocalizedthroughoutthetofurtherunravelingofthemechanismofplatinumw

7、holecell.After2±3hthecomplexaccumulatedinantitumorcomplexes.thenucleus,but6±8hafterincubationapunctuatestainingofacytoplasmicregionwasobserved,thatKeywordsCisplatin´Fluorescentplatinumpersistedandbecamemorepronouncedaf