ge11肽介导的靶向脂质体抗肿瘤作用及摄取机制的分析-analysis of the antitumor effect and uptake mechanism of targeted liposomes mediated by ge11 peptide.docx

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2018-06-04 发布于上海
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ge11肽介导的靶向脂质体抗肿瘤作用及摄取机制的分析-analysis of the antitumor effect and uptake mechanism of targeted liposomes mediated by ge11 peptide.docx

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ge11肽介导的靶向脂质体抗肿瘤作用及摄取机制的分析-analysis of the antitumor effect and uptake mechanism of targeted liposomes mediated by ge11 peptide

Studyoftheanti-tumoreffectandcellularuptakemechanismsofGE11peptidemodifiedtargetingliposomesAbstractObjective:Inthisstudy,weconstructedpeptideGE11decoratedDOX-loaded liposomesinordertoactivelytargettoEGFRover-expressedA549cells.Theinvitro cellexperimentswereusedtoevaluatethetargetingefficiency,cellularcytotoxicity, cellularuptake.We alsoevalued thetumoraccumulationofGE11 modified liposomes inA549tumor-bearingnudemice.Atlast,thecellularuptakemechanismofGE11 decoratedDOX-loadedliposomeswasstudied.Alloftheseexperimentsaimedto provide evidences forthis activetargetingdrugdeliverysystem against NSCLC.Methods:The DSPE-PEG2000-GE11were synthesized byadditionreaction betweenDSPE-PEG2000-MalandCys-GE11,thenthestructureoftheproductwas characterized by 1H-NMRandFTIRandtheconjugationefficiencyofGE11to DSPE-PEG2000-MalwasquantitativelydeterminedbyHPLC.TheGE11-LP/DOX waspreparedbycombinationofthethinfilmhydrationandpostinsertionmethod. Thentheparticlesize,zetapotential,morphologyandencapsulationefficiencyof liposomeswereevaluatedbyDLS,TEMandHPLC.TheMTTassaywasusedto choosetheoptimalGE11targetingdensityandevaluatetheIC50forfreeDOX, PEG-LP/DOXandGE11-LP/DOX.Qualitativeandquantitativeanalysisofliposomes cellularuptakebyA549cellswasdetectedbyfluorescencemicroscopeandflow cytometry.Thereal-timedistributionofliposomesinA549tumor-bearingnudemicewasdeterminedbyanear-infrared(NIR)fluorescenceimagingsystem.Atlast,the cellularuptakemechanismandthesubcellularlocalizationoftheliposomeswere examined byflowcytometryandconfocal microscopy.Results: The results of1H-NMR and FTIR verified the synthesis of theDSPE-PEG2000-GE11andthecalculatedconjugatedefficiencyofGE11to DSPE-PEG2000-Malwasabove90%detectedbyHPLC.TheparticlesizeoftheAbstractStudyoftheanti-tumor effectandcellular uptake mechanismsofGE11peptide modifiedtargetingliposomes differentGE11densitymodifiedliposomeswereabout124nm,thezetapotentialwas about?10mv.ThemorphologyofGE11-LP/DOXwasregularlysphericalinshape andtheencapsulationefficiencyo