Verotoxin sensitivity of ECV304 cells in vitro and in vivo in a xenograft tumour model: VT1 as a tumour neovascular marker.
Verotoxin sensitivity of ECV304 cells in vitro and in vivo in a xenograft tumour model: VT1 as a tumour neovascular marker.
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DOI：
10.1023/b:agen.0000011799.47529.fd
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发表时间：
2003-01-01
期刊：
影响因子：
9.8
通讯作者：
Lingwood, Clifford A.
中科院分区：
文献类型：
--
作者：
Heath-Engel, Hannah M.;Lingwood, Clifford A.
维罗毒素 1 (VT1) 通过其受体（globotriaosylceramide/Gb3）与内皮细胞和肿瘤细胞亚群结合，表明 VT1 可能具有抗肿瘤和抗血管生成的潜力。我们使用 ECV304 细胞系研究了这种潜力，该细胞系虽然被鉴定为膀胱癌细胞系，但显示出一些内皮特征，包括适当刺激后的小管形成（分化）。分化的 ECV304 细胞保留了 Gb3 表达/VT1 敏感性。观察到 VT1 内化和通过高尔基体逆行转运至内质网。免疫受损小鼠体内的 ECV304 异种移植物具有侵袭性，但令人惊讶的是血管化程度很差。瘤内注射 VT1 显着降低了 ECV304 异种移植物的生长并提高了小鼠的存活率。残余肿瘤中 Gb3 表达降低，可能是由于细胞周期停滞。未经处理的 ECV304 异种移植切片在整个肿瘤中结合 VT1。新血管系统的抗血管性血友病因子 (vWF) 抗体染色仅显示形态不典型、不清晰的结构，未用 VT1 标记。相比之下，人膀胱癌样本具有高度血管性，并且血管由抗 vWF 抗体和 VT1 100% 共同标记，没有血管外染色。这些结果表明，就 Gb3 表达而言，ECV304 异种移植物不是膀胱癌的特征，并且 VT1 染色可能提供肿瘤新生血管的新可靠指标。我们得出结论，ECV304 细胞不是肿瘤血管生成或膀胱癌的合适体内模型。然而，这些研究进一步证明了 VT1 的体内抗肿瘤和抗血管生成潜力，并表明 Gb3 在体外“血管”分化的细胞以及人类膀胱癌的新血管系统中表达。
Binding of verotoxin 1 (VT1) via its receptor (globotriaosylceramide/Gb3) to both endothelial and tumour cell subsets suggests that VT1 may have both antineoplastic and antiangiogenic potential. We investigated this potential using the ECV304 cell line, which, although identified as a bladder carcinoma cell line, displays some endothelial characteristics, including tubule formation (differentiation) following appropriate stimulation. Differentiated ECV304 cells retained Gb3 expression/VT1 sensitivity. VT1 internalization and retrograde transport through the Golgi to the ER was observed. ECV304 xenografts in immunocompromised mice were invasive but surprisingly poorly vascularized. Intratumoural VT1 injection significantly reduced ECV304 xenograft growth and enhanced mouse survival. Gb3 expression was decreased in residual tumour, likely due to cell cycle arrest. Untreated ECV304 xenograft sections bound VT1 throughout the tumour. Anti-von Willebrand factor (vWF) antibody staining for neovasculature showed only morphologically atypical, indistinct structures, unlabelled by VT1. Human bladder carcinoma samples were, in contrast, highly vascular and blood vessels were 100% co-labelled by anti-vWF antibody and VT1, with no extravascular staining. These results suggest that ECV304 xenografts are not characteristic of bladder carcinoma in terms of Gb3 expression, and that VT1 staining may provide a new reliable index of tumour neovasculature. We conclude that ECV304 cells are not an appropriate in vivo model of either tumour angiogenesis or bladder carcinoma. These studies, nevertheless, further demonstrate the in vivo antineoplastic and antiangiogenic potential of VT1, and show that Gb3 is expressed in cells undergoing in vitro 'vascular' differentiation, and in the neovasculature of human bladder carcinomas.