Pan Yunlong, Wu Qing, Qin Li, Cai Jiye, Du Bin
Department of General Surgery, The First Affiliated Hospital of Jinan University, Guangzhou 510632, China.
Department of Histology and Embryology, Medical School of Jinan University, Guangzhou 510632, China.
Biomed Res Int. 2014;2014:418624. doi: 10.1155/2014/418624. Epub 2014 Jun 1.
The early stages of angiogenesis can be divided into three steps: endothelial cell proliferation, migration, and tube formation. Vascular endothelial growth factor (VEGF) is considered the most important proangiogenic factor; in particular, VEGF165 plays a critical role in angiogenesis. Here, we evaluated whether gold nanoparticles (AuNPs) could inhibit the VEGF165-induced human umbilical vein endothelial cell (HUVEC) migration and tube formation. AuNPs and VEGF165 were coincubated overnight at 4°C, after which the effects on cell migration and tube formation were assessed. Cell migration was assessed using a modified wound-healing assay and a transwell chamber assay; tube formation was assessed using a capillary-like tube formation assay and a chick chorioallantoic membrane (CAM) assay. We additionally detected the cell surface morphology and ultrastructure using atomic force microscopy (AFM). Furthermore, Akt phosphorylation downstream of VEGFR-2/PI3K in HUVECs was determined in a Western blot analysis. Our study demonstrated that AuNPs significantly inhibited VEGF165-induced HUVEC migration and tube formation by affecting the cell surface ultrastructure, cytoskeleton and might have inhibited angiogenesis via the Akt pathway.
内皮细胞增殖、迁移和管腔形成。血管内皮生长因子(VEGF)被认为是最重要的促血管生成因子;特别是VEGF165在血管生成中起关键作用。在此,我们评估了金纳米颗粒(AuNPs)是否能够抑制VEGF165诱导的人脐静脉内皮细胞(HUVEC)迁移和管腔形成。将AuNPs与VEGF165在4°C下共同孵育过夜,之后评估其对细胞迁移和管腔形成的影响。使用改良的伤口愈合试验和Transwell小室试验评估细胞迁移;使用类毛细血管管腔形成试验和鸡胚绒毛尿囊膜(CAM)试验评估管腔形成。我们还使用原子力显微镜(AFM)检测了细胞表面形态和超微结构。此外,通过蛋白质印迹分析确定了HUVEC中VEGFR-2/PI3K下游的Akt磷酸化情况。我们的研究表明,AuNPs通过影响细胞表面超微结构、细胞骨架,显著抑制了VEGF165诱导的HUVEC迁移和管腔形成,并且可能通过Akt途径抑制了血管生成。