CAJ | 학술논문

目的探讨以VEGFR2 (kinase insert domain receptor,KDR)为靶点的靶向超声微泡对裸鼠结肠癌新生血管的成像效果.方法以生物素-亲和素桥接法将特异性结合VEGF主要受体KDR的小肽K237与脂质微泡耦联构建靶向微泡,用同样方法将对照肽与脂质微泡耦联,构建对照微泡.以KDR阴性表达的人结肠癌LS174T细胞株建立人结肠癌裸鼠移植瘤模型.12只荷瘤鼠经尾静脉随机先后注射靶向微泡、对照微泡,2种微泡注射间隔30 min.注射靶向微泡后5 min和注射对照微泡后5 min荷瘤鼠均行超声造影检查,观察各组微泡在肿瘤组织造影增强情况,测量肿瘤组织的声强度(Ⅵ).另取6只荷瘤鼠预先注射K237肽后再注射靶向微泡,观察微泡的成像效果.靶向微泡组、对照微泡组、小肽预先封闭组肿瘤组织的Ⅵ值比较采用单因素方差分析,组间多重比较采用最小显著性差异t检验.用免疫组织化学技术检测KDR在肿瘤组织表达及分布规律.结果成功制备了靶向微泡.注射超声微泡后5 min超声检查显示靶向微泡组肿瘤组织超声造影明显增强,对照微泡组及小肽预先封闭组仅见轻度的超声造影增强.3组Ⅵ值差异有统计学意义(F=39.130,P＜0.01).靶向微泡组与对照微泡组Ⅵ值差异有统计学意义(30.18±9.56与8.28±4.74,t=6.91,P＜0.01)；小肽预先封闭组Ⅵ值与靶向微泡组差异有统计学意义(9.23±3.44与30.18±9.56,t =4.91,P＜0.01).免疫组织化学结果显示,荷瘤鼠结肠癌新生血管内皮细胞KDR表达较正常组织血管内皮细胞KDR表达显著增加.结论以KDR为靶点的靶向超声微泡可以与荷瘤鼠肿瘤新生血管内皮特异性黏附并有效评价肿瘤新生血管形成.
목적 탐토이VEGFR2 (kinase insert domain receptor,KDR)위파점적파향초성미포대라서결장암신생혈관적성상효과.방법 이생물소-친화소교접법장특이성결합VEGF주요수체KDR적소태K237여지질미포우련구건파향미포,용동양방법장대조태여지질미포우련,구건대조미포.이KDR음성표체적인결장암LS174T세포주건립인결장암라서이식류모형.12지하류서경미정맥수궤선후주사파향미포、대조미포,2충미포주사간격30 min.주사파향미포후5 min화주사대조미포후5 min하류서균행초성조영검사,관찰각조미포재종류조직조영증강정황,측량종류조직적성강도(Ⅵ).령취6지하류서예선주사K237태후재주사파향미포,관찰미포적성상효과.파향미포조、대조미포조、소태예선봉폐조종류조직적Ⅵ치비교채용단인소방차분석,조간다중비교채용최소현저성차이t검험.용면역조직화학기술검측KDR재종류조직표체급분포규률.결과 성공제비료파향미포.주사초성미포후5 min초성검사현시파향미포조종류조직초성조영명현증강,대조미포조급소태예선봉폐조부견경도적초성조영증강.3조Ⅵ치차이유통계학의의(F=39.130,P＜0.01).파향미포조여대조미포조Ⅵ치차이유통계학의의(30.18±9.56여8.28±4.74,t=6.91,P＜0.01)；소태예선봉폐조Ⅵ치여파향미포조차이유통계학의의(9.23±3.44여30.18±9.56,t =4.91,P＜0.01).면역조직화학결과현시,하류서결장암신생혈관내피세포KDR표체교정상조직혈관내피세포KDR표체현저증가.결론 이KDR위파점적파향초성미포가이여하류서종류신생혈관내피특이성점부병유효평개종류신생혈관형성.
Objective To evaluate the effect of tumor neovascularization imaging in a nude mouse model of colon cancer by contrast ultrasound molecular imaging (UMI) of VEGF receptor 2 (kinase insert domain receptor,KDR).Methods Targeted microbubbles (MBt) were built by conjugating K237,a small peptide with high affinity for KDR,to liposome microbubbles through a biotin-avidin bridge.Control microbubbles (MBc) with control peptide were prepared by the same method.Nude mice models of LS174T human colon cancer were established.MBt and MBc were injected intravenously in twelve mice in random order with an interval of 30 min.MBt were injected in another six mice after K237-peptide blocking.UMI was performed in all mice at 5 min postinjection to observe the imaging difference and measure the video intensity (Ⅵ) of tumor tissues in different groups.One-way analysis of variance and the least significant difference t test were performed to analyze the difference of tumor VI in the groups with MBt,MBc and K237 blocking.Immunohistochemistry was applied to detect the expression and distribution of KDR in tumor tissue and adjacent tumor tissues.Results K237 peptide was successfully conjugated to the surface of microbubbles through biotin-avidin mediation.Ultrasound imaging signal of the tumor was high in the MBt group,while there were no significant enhancement in the groups of K237 blocking and MBc.The VI in MBt,MBc and K237 blocking groups was significantly different (F =39.130,P ＜ 0.01).There was a significant difference of VI in the MBt group compared to the MBc group (30.18 ± 9.56 vs 8.28 ± 4.74,t =6.91,P ＜0.01).In the K237 blocking group Ⅵ was significantly lower than that in the MBt group (9.23 ± 3.44 vs 30.18 ± 9.56,t =4.91,P ＜ 0.01).Immunohistochemistry results showed that KDR was highy expressed in tumor tissue.Conclusions KDR-targeting liposome contrast microbubbles may specifically and efficiently link to tumor vascular endothelial cells in vivo.Thus it may be used for UMI of tumor angiogenesis.