College of Bioengineering, Henan University of Technology, Zhengzhou 450001, China.
Acta Pharmacol Sin. 2013 Sep;34(9):1174-82. doi: 10.1038/aps.2013.60. Epub 2013 Jun 17.
Trans-3,4,5,4'-tetramethoxystilbene (DMU-212) has shown strong antiproliferative activities against a variety of cancer cells. The aim of this study was to investigate the anti-angiogenic effects of DMU-212 in vitro and in vivo.
Human umbilical vein endothelial cells (HUVECs) were used in this study. Cell viability was studied with MTT assay, and cell apoptosis was evaluated using TUNEL assay and morphological observation. The expression of the related genes and proteins was analyzed with qRT-PCR and Western blot, respectively. Angiogenesis of HUVECs were studied using cell migration and capillary-like tube formation assays in vitro, and mouse Matrigel plug assay and chick chorioallantoic membrane (CAM) assay in vivo. The tyrosine kinase activities of VEGFR1 and VEGFR2 were measured using commercial kits.
DMU-212 (5-80 μmol/L) significantly inhibited VEGF-stimulated proliferation of HUVECs (IC50 value was approximately 20 μmol/L), and induced apoptosis. Furthermore, DMU-212 concentration-dependently inhibited VEGF-induced migration of HUVECs and capillary-like structure formation in vitro. DMU-212 also inhibited VEGF-induced generation of new vasculature in Matrigel plugs in vivo with significantly decreased area of infiltrating CD31-positive endothelial cells, and inhibited newly formed microvessels in chick CAMs. Moreover, DMU-212 concentration-dependently suppressed VEGF-induced phosphorylation of VEGFR2, and inhibited phosphorylation of multiple downstream signaling components in the VEGFR2 pathway, including c-Src, FAK, Erk1/2, Akt, mTOR, and p70S6K in HUVECs. DMU-212 had no effect on VEGF-induced phosphorylation of VEGFR1.
DMU-212 is a potent inhibitor of angiogenesis that exerts anti-angiogenic activity at least in part through the VEGFR2 signaling pathway.
3,4,5,4'-四甲氧基二苯乙烯（DMU-212）对多种癌细胞表现出强烈的增殖抑制活性。本研究旨在探讨 DMU-212 的体外和体内抗血管生成作用。
本研究使用人脐静脉内皮细胞（HUVEC）。通过 MTT 测定法研究细胞活力，通过 TUNEL 测定法和形态观察评估细胞凋亡。分别使用 qRT-PCR 和 Western blot 分析相关基因和蛋白质的表达。在体外通过 HUVEC 细胞迁移和毛细血管样管形成测定法，以及体内通过小鼠 Matrigel plugs 测定法和鸡胚绒毛尿囊膜（CAM）测定法研究血管生成。使用商业试剂盒测量 VEGFR1 和 VEGFR2 的酪氨酸激酶活性。
DMU-212（5-80 μmol/L）显著抑制 VEGF 刺激的 HUVEC 增殖（IC50 值约为 20 μmol/L），并诱导细胞凋亡。此外，DMU-212 浓度依赖性地抑制 VEGF 诱导的 HUVEC 迁移和体外毛细血管样结构形成。DMU-212 还抑制 VEGF 诱导的 Matrigel plugs 中新血管生成，导致浸润的 CD31 阳性内皮细胞面积明显减少，并抑制鸡 CAM 中新生微血管形成。此外，DMU-212 浓度依赖性地抑制 VEGF 诱导的 VEGFR2 磷酸化，并抑制 VEGFR2 通路中的多个下游信号成分的磷酸化，包括 HUVEC 中的 c-Src、FAK、Erk1/2、Akt、mTOR 和 p70S6K。DMU-212 对 VEGF 诱导的 VEGFR1 磷酸化没有影响。
DMU-212 是一种有效的血管生成抑制剂，至少部分通过 VEGFR2 信号通路发挥抗血管生成活性。
