临床肿瘤学杂志

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重组人血管内皮抑素对血管新生的影响研究

叶庆1,2,3,秦叔逵2,殷晓进4,刘艳红3,丰俊东2,吴 穷2,曲文书2   

  1. 1 210002 南京 南京军区南京总医院博士后流动站2 210002 解放军八一医院全军肿瘤中心3 210008 南京大学医学院附属鼓楼医院病理科4 210042 江苏先声药物研发研究院
  • 收稿日期:2014-07-21 修回日期:2014-09-17 出版日期:2014-12-31 发布日期:2014-12-31
  • 通讯作者: 秦叔逵

Influences of recombinant human endostatin on angiogenesis

YE Qing, QIN Shukui, YIN Xiaojin, LIU Yanhong, FENG Jundong, WU Qiong, QU Wenshu.   

  1. Postdoctor Moving Station, General Hospital of Nanjing Military Command, Nanjing 210002, China
  • Received:2014-07-21 Revised:2014-09-17 Online:2014-12-31 Published:2014-12-31
  • Contact: QIN Shukui

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。

Abstract: 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 50000ng/ml, and its significant concentrations were 50ng/ml and 500ng/ml. Endostar inhibited HUVEC migration towards damage between 50ng/ml and 50000ng/ml in a dosedependent manner. Compared with 0ng/ml, the endostar at the dose of 50, 500 and 5000ng/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 50ng/ml and 50000ng/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.

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