Department of Urology, Daping Hospital, Army Medical University, Chongqing, People's Republic of China.
Nanotechnology. 2020 May 15;31(20):205101. doi: 10.1088/1361-6528/ab7040. Epub 2020 Feb 27.
Traditional imaging examinations have difficulty in identifying benign and malignant changes in renal masses. This difficulty may be solved by ultrasound molecular imaging based on targeted nanobubbles, which could specifically enhance the ultrasound imaging of renal cell carcinomas (RCC) so as to discriminate benign and malignant renal masses. In this study, we aimed to prepare anti-G250 nanobody-functionalized targeted nanobubbles (anti-G250 NTNs) by coupling anti-G250 nanobodies to lipid nanobubbles and to verify their target specificity and binding ability to RCC cells that express G250 antigen and their capacity to enhance ultrasound imaging of RCC xenografts. Anti-G250 nanobodies were coupled to the lipid nanobubbles using the biotin-streptavidin bridge method. The average particle diameter of the prepared anti-G250 NTNs was 446 nm. Immunofluorescence confirmed that anti-G250 nanobodies were uniformly distributed on the surfaces of nanobubbles. In vitro experiments showed that the anti-G250 NTNs specifically bound to G250-positive 786-O cells and HeLa cells with affinities of 88.13% ± 4.37% and 71.8% ± 5.7%, respectively, and that they did not bind to G250-negative ACHN cells. The anti-G250 NTNs could significantly enhance the ultrasound imaging of xenograft tumors arising from 786-O cells and HeLa cells compared with blank nanobubbles, while the enhancement was not significant for xenograft tumors arising from ACHN cells. Immunofluorescence of tumor tissue slices confirmed that the anti-G250 NTNs could enter the tissue space through tumor blood vessels and bind to tumor cells specifically. In conclusion, anti-G250 nanobody-functionalized targeted nanobubbles could specifically bind to G250-positive RCC cells and enhance the ultrasound imaging of G250-positive RCC xenografts. This study has high-potential clinical application value for the diagnosis and differential diagnosis of renal tumors.
传统的影像学检查在识别肾肿块的良恶性变化方面存在困难。这种困难可以通过基于靶向纳米气泡的超声分子成像来解决，它可以特异性地增强肾细胞癌 (RCC) 的超声成像，从而区分良性和恶性肾肿块。在这项研究中，我们旨在通过将抗 G250 纳米抗体偶联到脂质纳米气泡上来制备抗 G250 纳米抗体功能化的靶向纳米气泡（anti-G250 NTNs），并验证它们对表达 G250 抗原的 RCC 细胞的靶向特异性和结合能力，以及它们增强 RCC 异种移植瘤超声成像的能力。抗 G250 纳米抗体通过生物素-链霉亲和素桥接方法偶联到脂质纳米气泡上。所制备的抗 G250 NTNs 的平均粒径为 446nm。免疫荧光证实抗 G250 纳米抗体均匀分布在纳米气泡的表面上。体外实验表明，抗 G250 NTNs 特异性地与 G250 阳性的 786-O 细胞和 HeLa 细胞结合，亲和力分别为 88.13%±4.37%和 71.8%±5.7%，而与 G250 阴性的 ACHN 细胞不结合。与空白纳米气泡相比，抗 G250 NTNs 可显著增强源于 786-O 细胞和 HeLa 细胞的异种移植瘤的超声成像，而对源于 ACHN 细胞的异种移植瘤的增强作用不明显。肿瘤组织切片的免疫荧光证实，抗 G250 NTNs 可以通过肿瘤血管进入组织间隙并特异性地与肿瘤细胞结合。总之，抗 G250 纳米抗体功能化的靶向纳米气泡可以特异性地与 G250 阳性的 RCC 细胞结合，并增强 G250 阳性的 RCC 异种移植瘤的超声成像。这项研究对于肾肿瘤的诊断和鉴别诊断具有很高的临床应用价值。
