Establishment of canine hemangiosarcoma xenograft models expressing endothelial growth factors, their receptors, and angiogenesis-associated homeobox genes
Establishment of canine hemangiosarcoma xenograft models expressing endothelial growth factors, their receptors, and angiogenesis-associated homeobox genes
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DOI：
10.1186/1471-2407-9-363
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发表时间：
2009-10-14
期刊：
影响因子：
3.8
通讯作者：
Yanai, Tokuma
中科院分区：
文献类型：
--
作者：
Kodama, Atsushi;Sakai, Hiroki;Yanai, Tokuma
背景：人类血管肉瘤（HSA）往往预后不良；由于缺乏HSA临床标本，也没有HSA实验模型，其肿瘤发生机制尚未阐明。然而，犬类中自发性HSA的发生率相对较高；因此，犬HSA在人类HSA的研究中很有用。最近，已有报道在人和犬HSA中产生血管生长因子及其受体。此外，HSA的生长因子环境与病理生理性血管生成的生长因子环境非常相似，一些同源框基因在血管生成分子的转录中进行调节。在本研究中，我们建立了6个犬HSA异种移植瘤，并检测了生长因子及其受体和血管生成同源盒基因的表达。方法：将6个原代犬HSA异种移植到裸鼠皮下并连续移植。随后，通过组织病理学、免疫染色和反向研究，研究了血管内皮生长因子 (VEGF)-A、碱性成纤维细胞生长因子 (bFGF)、flt-1 和 flk-1（VEGF-A 受体）、FGFR-1 和血管生成同源盒基因 HoxA9、HoxB3、HoxB7、HoxD3、Pbx1 和 Meis1 在原始肿瘤和异种移植肿瘤中的表达。使用犬特异性引物组进行转录聚合酶链反应（RTPCR）。结果：从组织病理学角度来看，异种移植肿瘤包括形状各异的肿瘤细胞的增殖，从纺锤形、多边形到卵圆形；观察到一些血管样结构和通道的血管裂隙，与原始肿瘤相似。通过免疫组织化学和RT-PCR检测异种移植肿瘤中内皮标志物（CD31和vWF）的表达。此外，在异种移植肿瘤中检测到 VEGF-A、bFGF、flt-1、flk-1、FGFR-1、HoxA9、HoxB3、HoxB7、HoxD3、Pbx1 和 Meis1 的表达。有趣的是，3个HSA异种移植肿瘤中bFGF的表达往往高于其他肿瘤。结论：我们在裸鼠中建立了6个犬HSA异种移植肿瘤，发现异种移植HSA中血管生成因子及其受体的表达与自发HSA相似。此外，我们检测了血管生成同源盒基因的表达；因此，异种移植模型可能有助于分析 HSA 的恶性生长。
Background: Human hemangiosarcoma (HSA) tends to have a poor prognosis; its tumorigenesis has not been elucidated, as there is a dearth of HSA clinical specimens and no experimental model for HSA. However, the incidence of spontaneous HSA is relatively high in canines; therefore, canine HSA has been useful in the study of human HSA. Recently, the production of angiogenic growth factors and their receptors in human and canine HSA has been reported. Moreover, the growth-factor environment of HSA is very similar to that of pathophysiological angiogenesis, which some homeobox genes regulate in the transcription of angiogenic molecules. In the present study, we established 6 xenograft canine HSA tumors and detected the expression of growth factors, their receptors, and angiogenic homeobox genes.Methods: Six primary canine HSAs were xenografted to nude mice subcutaneously and serially transplanted. Subsequently, the expressions of vascular endothelial growth factor (VEGF)-A, basic fibroblast growth factors (bFGF), flt-1 and flk-1 (receptors of VEGF-A), FGFR-1, and angiogenic homeobox genes HoxA9, HoxB3, HoxB7, HoxD3, Pbx1, and Meis1 were investigated in original and xenograft tumors by histopathology, immunostaining, and reverse transcription polymerase chain reaction (RTPCR), using canine-specific primer sets.Results: Histopathologically, xenograft tumors comprised a proliferation of neoplastic cells that were varied in shape, from spindle-shaped and polygonal to ovoid; some vascular-like structures and vascular clefts of channels were observed, similar to those in the original tumors. The expression of endothelial markers (CD31 and vWF) was detected in xenograft tumors by immunohistochemistry and RT-PCR. Moreover, the expression of VEGF-A, bFGF, flt-1, flk-1, FGFR-1, HoxA9, HoxB3, HoxB7, HoxD3, Pbx1, and Meis1 was detected in xenograft tumors. Interestingly, expressions of bFGF tended to be higher in 3 of the xenograft HSA tumors than in the other tumors.Conclusion: We established 6 xenograft canine HSA tumors in nude mice and found that the expressions of angiogenic growth factors and their receptors in xenograft HSAs were similar to those in spontaneous HSA. Furthermore, we detected the expression of angiogenic homeobox genes; therefore, xenograft models may be useful in analyzing malignant growth in HSA.