RGD-Targeted Ultrasound Contrast Agent for Longitudinal Assessment of Hep-2 Tumor Angiogenesis In Vivo.
RGD-Targeted Ultrasound Contrast Agent for Longitudinal Assessment of Hep-2 Tumor Angiogenesis In Vivo.
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RGD 靶向超声造影剂用于体内 Hep-2 肿瘤血管生成的纵向评估。
DOI：
10.1371/journal.pone.0149075
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Wen ZH
中科院分区：
文献类型：
--
作者：
Hu Q;Wang XY;Kang LK;Wei HM;Xu CM;Wang T;Wen ZH
目的制备精氨酸-甘氨酸-天冬氨酸（RGD）靶向超声造影剂微泡（MBs），并探讨其用于评价肿瘤血管生成模型中αvβ3整合素表达动态变化的可行性。通过生物素-亲和素连接将RGD肽缀合至微泡表面。制备携带RADfK肽的微泡作为对照。使用Accusizer 780和光学显微镜表征RGD-MB。通过竞争性抑制实验在体外证明RGD-MB对表达ανβ3的内皮细胞（bEnd.3）的结合特异性。在一项体内研究中，对携带三个不同阶段肿瘤的小鼠静脉注射RGD-MB，并对其进行靶向、对比增强、高频超声。随后，收获肿瘤并切片用于ανβ3表达的免疫荧光分析。RGD-MB的平均大小为2.36 ± 1.7 μm。RGD-MB与bEnd.3细胞的粘附水平显著高于对照MB（P < 0.01）。RGD-MB很少与αvβ3阴性MCF-7细胞结合。用抗α（v）抗体处理后，RGD-MB与bEnd.3细胞的粘附受到显著抑制。高频超声造影对人喉癌（Hep-2）皮下移植瘤的定量声像图强度，小肿瘤（19.89 ± 2.49）明显高于中等肿瘤（11.25 ± 2.23）和大肿瘤（3.38 ± 0.67）（P < 0.01）。RGD-MB能够在皮下癌异种移植物中肿瘤生长期间对肿瘤血管生成的变化进行非侵入性体内可视化。
To prepare arginine-glycine-aspartate (RGD)-targeted ultrasound contrast microbubbles (MBs) and explore the feasibility of their use in assessing dynamic changes in αvβ3 integrin expression in a murine model of tumor angiogenesis. RGD peptides were conjugated to the surfaces of microbubbles via biotin-avidin linkage. Microbubbles bearing RADfK peptides were prepared as controls. The RGD-MBs were characterized using an Accusizer 780 and optical microscopy. The binding specificity of the RGD-MBs for ανβ3-expressing endothelial cells (bEnd.3) was demonstrated in vitro by a competitive inhibition experiment. In an in vivo study, mice bearing tumors of three different stages were intravenously injected with RGD-MBs and subjected to targeted, contrast-enhanced, high-frequency ultrasound. Subsequently, tumors were harvested and sectioned for immunofluorescence analysis of ανβ3 expression. The mean size of the RGD-MBs was 2.36 ± 1.7 μm. The RGD-MBs showed significantly higher adhesion levels to bEnd.3 cells compared to control MBs (P < 0.01). There was rarely binding of RGD-MBs to αvβ3-negative MCF-7 cells. Adhesion of the RGD-MBs to the bEnd.3 cells was significantly inhibited following treatment with anti-alpha(v) antibodies. The quantitative acoustic video intensity for high-frequency, contrast-enhanced ultrasound imaging of subcutaneous human laryngeal carcinoma (Hep-2) tumor xenografts was significantly higher in small tumors (19.89 ± 2.49) than in medium tumors (11.25 ± 2.23) and large tumors (3.38 ± 0.67) (P < 0.01). RGD-MBs enable noninvasive in vivo visualization of changes in tumor angiogenesis during tumor growth in subcutaneous cancer xenografts.