人离体骨肉瘤组织血管噬菌体肽库筛选模型的建立及其意义

Establishment of biopanning model of phage display peptide library in the blood vessels of excised human osteosarcoma vasculature and its significance

目的建立人离体骨肉瘤血管亲和筛选模型。方法(1)建立模型。术前对患肢局部行数字减影造影检查,标记肿瘤固有动脉大致走行,术后小心修剪,并连接类似于Langendorff的灌流装置,监控模型血管内的pH值、温度、氧分压等;(2)对噬菌体十二肽库进行筛选。结果28例骨肉瘤离体标本均建模成功,可直接用于灌流实验,并筛选出具有较高特异性的亲和短肽(基序为RLTR),各项指标显示离体骨肉瘤较好模拟了人体内的环境。结论建立人离体骨肉瘤血管亲和筛选模型是可行的,可直接用于对噬菌体随机肽库的亲和筛选,为骨肉瘤的靶向化疗提供参考。

Objective To establishan in vivo biopanningmodelof phagedisplaypeptidelibraryin the bloodvessels containedin surgicallyremovedhumanosteosarcoma.Method In28patientswithosteosarcoma,digitalsubtraction angiography(DSA)of theinvolvedlimbwas performedpreoperativelyto understandtheapproximatestatusof thearteriesin thetumors.Thetumorswerethensurgicallyremovedandcarefullytrimmed,perfusedviaa simulatedLangendorffperfusion apparatuswiththeindexesas thepHvalue,temperatureandO 2 partialpressuremonitoredinthebloodvessels.A12-meres phagedisplaypeptidelibrarywas biopannedto isolatepeptidescapableof homingspecificallyto theexcisedosteosarcoma.Results In vivo biopanningmodelsweresuccessfullyestablishedin alltheexcisedtumors,whichcouldbe directlyusedin perfusionexperimentandtheindexesmonitoredinthebloodvesselsinthetumorswerecomparableto thoseof livingtissues.Somehigh-affinitypeptidesspecificto thebloodvesselsinosteosarcomawereobtainedwiththemotifof RLTR.Conclusion In vivo biopanningmodelsimulatingtheLangendorffperfusionapparatuscanbe easilyestablishedwhichis instrumentalfor theapplicationof phagedisplaytechnologyin humanlivingtissuesandmayfacilitatethestudyof targetedchemotherapyof osteosarcoma.