摘要
目的探讨重组人血管内皮抑素(Endostar,恩度)对血管内皮细胞趋化、迁移、粘附、增殖及小管形成等与血管新生相关生物学行为的影响。方法以原代培养的人脐静脉内皮细胞(HUVEC)为细胞模型,通过Boyden小室荧光定量分析、划痕试验、HUVEC荧光定量粘附分析、CFSE染色流式细胞术、CCK-8定量检测、小管形成试验和Matrigel栓试验研究恩度对HUVEC与血管新生相关的生物学行为的影响。结果恩度在5~50 000ng/ml范围内可抑制血管内皮生长因子诱导HUVEC的迁移运动,且浓度为50ng/ml和500ng/ml时效果最明显;恩度在5~50 000ng/ml间可呈剂量依赖的方式抑制HUVEC向损伤部位的迁移。与0ng/ml相比,50、500和5000ng/ml恩度处理的HUVEC的粘附率、增殖率及HUVEC形成网状小管结构的数量、面积和长度均降低,差异均有统计学意义(P〈0.05);Matrigel栓实验结果显示,恩度在50~5000ng/ml间对SCID小鼠体内血管新生有明显的抑制作用。结论恩度在细胞水平能抑制HUVEC与血管新生相关的生物学行为,包括HUVEC的趋化、迁移、粘附、增殖和小管形成;在动物水平能抑制SCID小鼠体内的血管新生,据此推断恩度能抑制血管新生。
Objective To study the effects of recombinant human endostatin( endostar) on the angiogenesis-related biological behaviors such as chemotaxis,migration,adhesion,proliferation and tube formation of vascular endothelial cells. Methods With primary cultured human umbilical vein endothelial cells( HUVEC) as a model,the fluorescence quantitative Boyden chamber analysis,wound healing assay,fluorescence quantitative adhesion assay,flow cytometry,tube formation assay and Matrigel plug test were employed to evaluate the effects of endostar on the angiogenesis-related biological behavior of HUVECs in vitro and in vivo. Results Endostar potently inhibited HUVEC migration in response of VEGF from 50 to 50000 ng / ml,and its significant concentrations were 50 ng /ml and 500 ng / ml. Endostar inhibited HUVEC migration towards damage between 50 ng / ml and 50000 ng / ml in a dose-dependent manner. Compared with 0ng / ml,the endostar at the dose of 50,500 and 5000 ng / ml decreased adhesion rate and proliferation rate of HUVEC cells as well as the number,area and length of HUVEC mesh tubular with statistically significant difference( P 〈 0. 05). Matrigel plug test also revealed that Endostar inhibited Matrigel plug formation in SCID mice between 50 ng / ml and 50000 ng / ml. Conclusion At the cellular level,endostar inhibited the angiogenesis-related biological behaviors of HUVECs,including migration,adhesion,proliferation and tube formation. In vivo,endostar inhibited the angiogenesis in SCID mice.
出处
《临床肿瘤学杂志》
CAS
2014年第12期1062-1068,共7页
Chinese Clinical Oncology