Chen Ya-Huey, Wu Hua-Lin, Li Ching, Huang Yi-Hsien, Chiang Chi-Wu, Wu Ming-Ping, Wu Li-Wha
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, 1 University Road, Tainan 70101, Taiwan, Republic of China.
Thromb Haemost. 2006 Apr;95(4):668-77.
The molecular mechanism mediated by multiple forms of angiostatin via acting on proliferating vascular endothelium remains elusive. To address whether three forms of angiostatin, K1-3, K1-4 or K1-4.5, utilized similar or distinct pathways to mediate anti-angiogenesis, we adopted an adenoviral expression system to express secretable angiostatin molecules for CM collection. The anti-angiogenic activity of K1-3, K1-4 or K1-4.5 was confirmed by using proliferation, migration, tube formation and apoptotic assays of human endothelial cells. These angiostatin molecules at comparable expression level inhibited various in vitro angiogenesis assays with some variations. Furthermore, K1-3, K1-4 or K1-4.5 increased the expression of p53 protein and its downstream effectors, enhanced FasL-mediated signaling pathways, and decreased activation of AKT. At least three different receptors, Fas, integrin alpha(v)beta3 and ATP synthase, were involved in the anti-angiogenic action of angiostatin molecules. Besides, the expression of 189 genes at mRNA level was significantly altered by K1-3, K1-4 or K1-4.5. More than 70% of these genes participate in growth, inflammation, apoptosis, migration and extracellular matrix. Taken together, K1-3, K1-4 and K1-4.5, regardless of the number of kringles in the angiostatin molecules, mediated anti-angiogenesis via mostly similar pathways. We are the first to demonstrate the involvement of DAPK1 in the mediation of anti-angiogenesis by angiostatin.
多种形式的血管抑素通过作用于增殖的血管内皮细胞所介导的分子机制仍不清楚。为了探究血管抑素的三种形式K1-3、K1-4或K1-4.5是否利用相似或不同的途径来介导抗血管生成作用，我们采用腺病毒表达系统来表达可分泌的血管抑素分子以收集条件培养基。通过对人内皮细胞进行增殖、迁移、管腔形成和凋亡检测，证实了K1-3、K1-4或K1-4.5的抗血管生成活性。这些表达水平相当的血管抑素分子在体外抑制各种血管生成检测时存在一些差异。此外，K1-3、K1-4或K1-4.5增加了p53蛋白及其下游效应分子的表达，增强了FasL介导的信号通路，并降低了AKT的活性。血管抑素分子的抗血管生成作用至少涉及三种不同的受体，即Fas、整合素α(v)β3和ATP合酶。此外，K1-3、K1-4或K1-4.5显著改变了189个基因的mRNA水平表达。其中超过70%的基因参与生长、炎症、凋亡、迁移和细胞外基质过程。综上所述，无论血管抑素分子中kringle结构域的数量如何，K1-3、K1-4和K1-4.5大多通过相似的途径介导抗血管生成作用。我们首次证明DAPK1参与血管抑素介导的抗血管生成作用。
