CAJ | 학술논문

目的验证骨肉瘤血管内皮细胞(osteosarcoma-associated vascular endothelial cells,OAVECs)特异性结合肽(TKPDKGY)对荷瘤裸鼠模型体内OAVECs的亲和性.方法合成短肽(TKPD-KGY)并在其N端加载一个异硫氰酸荧光素(fluorescein isothiocyanate,FITC),纯化后将短肽荧光复合物注入荷瘤裸鼠血管内,通过小动物整体荧光成像来观察短肽在活体动物内的分布,同时定时处死裸鼠,切取重要器官和肿瘤制冰冻切片,使用激光共聚焦显微镜扫描来评价短肽在体内与OAVECs的亲和性.结果小动物整体荧光成像显示荷瘤裸鼠注射FTTC-TKPDKGY 10 min后,肿瘤处即可见微弱荧光富集,30min时显示明显并逐渐增强,1h后达最强,4 h后逐渐消退,至24 h基本无荧光残留.激光共聚焦扫描可见肿瘤血管和血管丛荧光强度较强,肿瘤组织仅见少量短肽结合,可提示有一定的结合性.心、脑、肺和肝组织显示并不明显.肾脏镜下可见肾血管并无荧光显像,但肾小球和肾小管部可见荧光富集,提示短肽在此代谢.结论 TKPDKGY对目标细胞有良好的靶向性,为短肽进一步结合效应分子应用于骨肉瘤的诊断和治疗奠定一定的实验基础.
목적 험증골육류혈관내피세포(osteosarcoma-associated vascular endothelial cells,OAVECs)특이성결합태(TKPDKGY)대하류라서모형체내OAVECs적친화성.방법 합성단태(TKPD-KGY)병재기N단가재일개이류청산형광소(fluorescein isothiocyanate,FITC),순화후장단태형광복합물주입하류라서혈관내,통과소동물정체형광성상래관찰단태재활체동물내적분포,동시정시처사라서,절취중요기관화종류제빙동절편,사용격광공취초현미경소묘래평개단태재체내여OAVECs적친화성.결과 소동물정체형광성상현시하류라서주사FTTC-TKPDKGY 10 min후,종류처즉가견미약형광부집,30min시현시명현병축점증강,1h후체최강,4 h후축점소퇴,지24 h기본무형광잔류.격광공취초소묘가견종류혈관화혈관총형광강도교강,종류조직부견소량단태결합,가제시유일정적결합성.심、뇌、폐화간조직현시병불명현.신장경하가견신혈관병무형광현상,단신소구화신소관부가견형광부집,제시단태재차대사.결론 TKPDKGY대목표세포유량호적파향성,위단태진일보결합효응분자응용우골육류적진단화치료전정일정적실험기출.
Objective To identify the binging specification of the short peptide(TKPDKGY)specifi-cally binding to osteosarcoma-associated vascular endothelial cells(OAVECs)by using BALB/c-nu mice with osteosarcoma.Methods The TKPDKGY was synthesized with the N-terminal marked with fluorescein isoth-iocyanate(FITC).After purification,the peptide was injected into the vessel of nude mice.Whole-body optical imaging system was used to investigate the peptide distribution in mice.Four hours after injection,the nude mice were sacrificed.The vital organs and tumor of mice were used to make frozen slice.Confocal micro-scope was used to detect the in-vivo binding activity of the short peptide.Results Whole-body optical imaging system showed that fluorescence at tumor site was weak at 10 min after injection,and gradually in-creased at 30 min,reached the peak at 1 h,then subsidised 4 h later,and no residual fluorescence at 24 h.Confocal microscope also to shown that the short peptide homed to vascular endothelium in osteosarcoma but was not detectable in the heart,brain,lung and liver.The fluorescence at glomerular and tubular was detect-ed by confocal microscope but not renal vascular endothelium which means that the kidney is metabolism or-gan of TKPDKGY.Conclusion The TKPDKGY peptide has the specifically binding activity to vascular en-dothelium in osteosarcoma.It may be used as good targeting vector for selective delivery of therapeutics and as a diagnostic probe for the detection of osteosarcoma.