Antiangiogenic activity of genistein in pancreatic carcinoma cells is mediated by the inhibition of hypoxia-inducible factor-1 and the down-regulation of VEGF gene expression
Antiangiogenic activity of genistein in pancreatic carcinoma cells is mediated by the inhibition of hypoxia-inducible factor-1 and the down-regulation of VEGF gene expression
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DOI：
10.1002/cncr.11873
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发表时间：
2004-01-01
期刊：
影响因子：
6.2
通讯作者：
Hines, OJ
中科院分区：
文献类型：
--
作者：
Büchler, P;Reber, HA;Hines, OJ
背景。以前的报道表明染料木素是一种天然存在的异黄酮，具有很强的抗血管生成活性。然而，抑制的潜在机制尚不清楚。染料木素的生物学作用包括抑制酪氨酸激酶和抑制缺氧诱导因子-1 （HIF-1）的缺氧激活，HIF-1是VEGF基因表达的主要调节因子之一。在模块化培养箱中进行缺氧细胞培养。采用酶联免疫吸附法测定血管内皮生长因子（VEGF）蛋白的分泌，电泳迁移位移法测定HIF-1对DNA的结合，Northern blot法测定mRNA的含量。采用原位小鼠胰腺癌模型进行研究。通过对异种移植肿瘤的内皮细胞标记物进行染色，定量研究了体内血管生成。随着染料木黄酮剂量的增加，VEGF蛋白分泌呈剂量依赖性抑制。此外，用染料木黄酮处理胰腺癌细胞导致缺氧培养条件下HIF-1的激活受损。Northern blot分析显示，在常氧和缺氧培养条件下，染料木素处理后VEGF mRNA表达均下降。在体内，染料木素抑制异种移植胰腺癌细胞的肿瘤生长，而在异种移植肿瘤中观察到广泛的缺氧，并且不受染料木素治疗的影响。同样，在染料木黄酮处理的Capan-1异种移植肿瘤中，VEGF mRNA水平下降。目前的研究表明，以前报道的染料木素的抗血管生成活性可能是通过抑制HIF-1介导的，HIF-1是VEGF基因稳态的重要调节因子，特别是在低氧条件下。因此，这种生物活性化合物可能对胰腺癌患者有益。(C) 2003年美国癌症协会。
BACKGROUND. Previous reports indicate that Genistein, a naturally occurring isoflavonoid, exhibits strong antiangiogenic activity. The underlying mechanism of inhibition, however, remains unclear. Among the biologic effects of Genistein are the inhibition of tyrosine kinases and the inhibition of hypoxic activation of hypoxia-inducible factor-1 (HIF-1), one of the main regulators of VEGF gene expression.METHODS. Hypoxic cell culture was performed in a modular incubator chamber. Vascular endothelial growth factor (VEGF) protein secretion was measured using the enzyme-linked immunosorbent assay, binding of DNA by HIF-1 was measured using the electrophoretic mobility shift assay, and mRNA quantification was performed using Northern blot analysis. Pancreatic carcinoma was studied in an orthotopic murine model. Angiogenesis in vivo was quantified by staining xenograft tumors for endothelial cell markers.RESULTS. VEGF protein secretion was dose-dependently suppressed with increasing doses of Genistein. Furthermore, treatment of pancreatic carcinoma cells with Genistein led to impaired activation of HIF-1 under hypoxic culture conditions. Northern blot analysis indicated that VEGF mRNA expression decreased upon treatment with Genistein, both under normoxic and hypoxic culture conditions. In vivo, Genistein inhibited tumor growth for xenograft pancreatic carcinoma cells, whereas extensive hypoxia was observed in xenograft tumors and was not influenced by Genistein therapy. Similarly, decreased VEGF mRNA levels were observed in Genistein-treated Capan-1 xenograft tumors.CONCLUSIONS. The current study indicates that the previously reported antiangiogenic activity of Genistein probably is mediated by the inhibition of HIF-1, an important regulator of VEGF gene homeostasis, particularly under low-oxygen conditions. Therefore, this bioactive compound may well be beneficial to patients with pancreatic carcinoma. (C) 2003 American Cancer Society.