双靶超声微泡评估肾癌生长过程中促血管生成因子表达水平的实验研究  

Experimental study of dual-targeted microbubbles in assessing the expression levels of pro-angiogenic markers during the growth of renal carcinoma

作　　者：李翠仙 黄备建 陆清 陆蓓蕾 李丛 王文平 Li Cuixian;Huang Beijian;Lu Qing;Lu Beilei;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12期1077-1083,共7页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金(81901750,12174074);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(VEGFR)/整合素α_(v)β_(3)双靶超声造影剂评估肾细胞癌(RCC)生长过程中肿瘤促血管生成因子表达水平的能力。方法:采用生物素-亲和素桥接法制备VEGFR/整合素α_(v)β_(3)双靶超声造影剂。构建20只人肾脏透明细胞癌(786-O细胞株)裸鼠皮下种植瘤模型并将其随机分为2组。第一组用于纵向评价RCC生长过程中靶向超声造影定量参数变化情况,该组10只荷瘤鼠均接受2次靶向超声造影检查,2次检查时瘤体最大径分别为5~10 mm、>10~20 mm。第二组分为A、B两个亚组,每组5只荷瘤鼠:A组裸鼠瘤体最大径为5~10 mm,B组最大径为>10~20 mm,荷瘤鼠均行靶向超声造影检查。对超声造影声像图行定量分析后获取以下靶向定量参数:造影前3 min感兴趣区峰值强度(PI)、曲线下面积(AUC),超声爆破前及爆破后10 s各自峰值强度P 1及P 2,以及P 1与P 2的差值(dTE)。检查结束后获取第二组荷瘤鼠肿瘤组织行免疫组化染色,观察其内促血管生成因子VEGFR2、整合素α_(v)β_(3)及CD31表达情况,以横向比较各因子在不同大小RCC中的表达差异,并分析靶向载药定量参数与各因子表达量间相关性。结果:纵向分析结果显示,不同大小RCC靶向定量参数AUC及dTE差异有统计学意义(均P<0.001),瘤体越大,靶向定量参数越小。横向对比结果显示,不同大小RCC间VEGFR2及整合素α_(v)β_(3)表达水平差异有统计学意义(均P<0.05),CD31表达水平差异则无统计学意义(P=0.754)。此外,瘤体最大径与靶向定量参数AUC、dTE、VEGFR2、整合素α_(v)β_(3)水平呈显著负相关(r_(s)=-0.83、-0.81、-0.70、-0.88,均P<0.05),AUC与VEGFR2、整合素α_(v)β_(3)表达水平间呈显著正相关(r_(s)=0.76、0.72,均P<0.05),dTE与VEGFR2及整合素α_(v)β_(3)表达水平间呈显著正相关(r_(s)=0.81、0.72,均P<0.05),PI与CD31表达水平呈正相关(r_(s)=0.70,P=0.025)。结论:RCC内促血管生成因子VEGFR2及整Objective To explore the capability of vascular endothelial growth factor receptor 2(VEGFR2)/integrinα_(v)β_(3) dual-targeted microbubbles in assessing the expression level of pro-angiogenic factors during renal cell carcinoma(RCC)growth.Methods VEGFR2/integrinα_(v)β_(3) dual-targeted microbubbles were prepared by using biotin-avidin linkage method.Twenty subcutaneous RCC xenografts in nude mice were established by subcutaneously injecting 786-O cells and then divided into 2 groups randomly.The targeted contrast-enhanced ultrasound(t-CEUS)examination was performed for all 10 mice in the first group when xenograft tumors were metered from 5 to 10 mm and>10 to 20 mm respectively.And the quantitative parameters of RCC on t-CEUS were longitudinally evaluated during tumor growth.The second group were divided into two subgroups according to xenograft tumors′diameter,which was 5 to 10 mm and>10 to 20 mm respectively,and underwent t-CEUS examination.Quantitative analysis was performed for all t-CEUS images to obtain the targeted quantitative parameters,which including peak intensity(PI),area under the time-intensity curve(AUC),the differential tissue enhancement(dTE,presenting the difference in PI before(P1)and after(P2)the process of Flash).All xenograft tumors in the second group were harvested for immunohistochemical staining to observe the expression of VEGFR2,integrinα_(v)β_(3) and CD31,and their differences in RCC with different tumor sizes.And the correlations between quantitative parameters and VEGFR2,integrinα_(v)β_(3) and CD31 were analyzed.Results The longitudinal comparison showed that there were statistically significant differences between AUC and dTE of RCC with different tumor sizes(all P<0.001).The larger the tumor size,the smaller the parameters were.According to the horizontal comparison,the expression levels of VEGFR2 and integrinα_(v)β_(3) in larger RCCs were higher than those of RCCs with smaller size(both P<0.05),but there was no significant difference in CD31 expression between the two

关 键 词：超声造影 靶向微泡 肾细胞癌 血管生成 

分 类 号：R737.11[医药卫生—肿瘤] R445.1[医药卫生—临床医学]