靶向血管内皮生长因子受体2/整合素α_(v)β_(3)微泡体内评价肾细胞癌血管生成的实验研究  

Experimental study of vascular endothelial growth factor receptor 2 and integrinα_(v)β_(3) dual-targeted microbubble to assess tumor angiogenesis of renal cell carcinoma in vivo

作　　者：李翠仙 黄备建 陆清 陆蓓蕾 袁海霞 李丛 王文平 Li Cuixian;Huang Beijian;Lu Qing;Lu Beilei;Yuan Haixia;Li Cong;Wang Wenping(Department of Ultrasound,Zhongshan Hospital of Fudan University,Shanghai 200032,China;Shanghai Institute of Imaging Medicine,Shanghai 200032,China)

机构地区：[1]复旦大学附属中山医院超声科,上海200032 [2]上海市影像医学研究所,上海200032

出　　处：《中华超声影像学杂志》2022年第4期338-344,共7页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金项目(81901750);上海市卫计委临床专项研究(青年项目)(20184Y0366);上海市临床重点专科(shslczdzk03501);上海市"医苑新星"青年医学人才项目(R2021-007)。

摘　　要：目的探讨血管内皮生长因子受体2(VEGFR2)/整合素α_(v)β_(3)双靶向造影剂(MBD)在体内对肾细胞癌肿瘤新生血管的靶向能力。方法以造影剂USphere LA为模板,采用生物素-亲和素桥接法制备以VEGFR2、整合素α_(v)β_(3)为靶点的单靶向造影剂及双靶向造影剂。构建人肾脏透明细胞癌(786-O细胞株)裸鼠皮下种植瘤模型共40只,选取其中20只,每只均以任意顺序采用4种造影剂[非靶向造影剂(MBN)、VEGFR2单靶向造影剂(MBV)或整合素α_(v)β_(3)单靶向造影剂(MBI)及MBD]进行超声造影检查;另20只裸鼠行抗体阻断试验。所得声像图进行定量分析,得到以下定量参数:造影前3 min造影剂强度增量(a_(1))、峰值减半速度(a_(2))、曲线上升斜率(a_(3))、灌注时间(t_(0))、达峰时间(TTP)、峰值强度(PI)、平均渡越时间(MTT)和ROC曲线下面积(AUC),爆破前及爆破后10 s的峰值强度(P1及P2),以及二者的差值(dTE),比较4种造影剂间及抗体阻断前后各定量参数在不同组间的差异。免疫组化染色观察肿瘤组织CD31、VEGFR2及整合素β3表达情况。结果将MBN、MBV或MBI及MBD进行比较,结果显示2种单靶向造影剂间所有参数差异无统计学意义(均P>0.05),a_(1)、a_(3)、t_(0)、TTP、PI及P2在4种造影剂间差异无统计学意义(均P>0.05);AUC、MTT、P1及dTE表现为双靶向造影剂>单靶向造影剂>非靶向造影剂的趋势(均P<0.05),与之相反,参数a_(2)则表现为双靶向造影剂<单靶向造影剂<非靶向造影剂的趋势(均P<0.05)。比较双靶向造影剂MBD各定量参数在抗体阻断前后的差别显示,抗体阻断后a_(2)快于抗体阻断前(P<0.001),而AUC、MTT、P1及dTE较抗体阻断前降低(均P<0.001),其余参数在抗体阻断前后差异无统计学意义(均P>0.05)。免疫组化染色结果显示人肾细胞癌组织内有明显的CD31、VEGFR2及整合素β3表达。结论以VEGFR2及整合素α_(v)β_(3)为靶点的双靶向造影剂在体内对人肾细胞�Objective To evaluate the ability of vascular endothelial growth factor receptor 2(VEGFR2)/integrinα_(v)β_(3) dual-targeted microubble(MBD)to target angiogenesis of renal cell carcinoma(RCC)in vivo.Methods Non-targeted microbubble(MBN)USphere LA was employed as a template to prepare single-and dual-targeted microbubbles which could bind VEGFR2 and/or integrinα_(v)β_(3)(MBV and MBI)by the biotin-avidin bridging method.A total of 40 RCC nude mice models were established by subcutaneously injecting 786-O cells.Twenty of the models were all injected with MBN,MBV,MBI and MBD in a random order,and the other 20 models were registered for antibody blocking assays.The results of ultrasound images were used for quantitative analyses,and the following quantitative parameters were obtained:intensity increment(a_(1)),peak halving speed(a_(2)),curve rising slope(a_(3)),perfusion time(t_(0)),time to peak(TTP),peak intensity(PI),mean transit time(MTT)and area under the curve(AUC)for the first three minutes,peak intensity at 10 s before(P1)and after(P2)ultrasound destruction,and the differences of tissue enhancement(dTE)between P1 and P2(dTE=P1-P2).All the quantitative parameters of four contrast agents and the antibody blocking assays were compared.Besides,the immunohistochemical assays were performed to evaluate the expression of CD31,VEGFR2 and integrinα_(v)β_(3) in tumor tissues.Results The differences of parameters of a_(1),a_(3),t_(0),TTP,PI and P2 among four different microbubbles had no statistical significances(all P>0.05),and all parameters between the two single-targeted contrast agents were not statistically different(all P>0.05).The parameters of AUC,MTT,P1 and dTE all showed a trend that dual-targeted bubbles>single-targeted bubbles>non-targeted bubbles(all P<0.05).On the contrary,the trend of dual-targeted bubbles<single-targeted bubbles<non-targeted bubbles(all P<0.05)was observed for a_(2).In the antibody blocking experiment,a_(2) was faster after the antibody injection(P<0.001),while AUC,MTT,P1 and dTE wer

关 键 词：超声造影 靶向微泡 肾细胞癌 血管生成 血管内皮生长因子受体2 整合素α_(v)β_(3) 

分 类 号：R737.11[医药卫生—肿瘤]