血管内皮生长因子（VEGF）与胸苷磷酸化酶（TP）联合应用以改善血管生成基因治疗。

combination of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP) to improve angiogenic gene therapy.

作者信息

Bouïs Diane, Boelens Mirjam C, Peters Erna, Koolwijk Pieter, Stob Gerrie, Kema Ido P, Klinkenberg Marco, Mulder Nanno H, Hospers Geke A P

机构信息

Department of Medical Oncology, University Hospital Groningen, The Netherlands.

出版信息

Angiogenesis. 2003;6(3):185-92. doi: 10.1023/B:AGEN.0000021389.49659.31.

DOI:10.1023/B:AGEN.0000021389.49659.31

PMID:15041794

Abstract

To improve current angiogenic gene therapy with a vascular endothelial growth factor (VEGF)-encoding plasmid (Baumgartner et al. Circulation 1998; 97: 1114-23 [1]; Kusumanto et al. Fifth Annual Meeting of the American Society of Gene Therapy, Boston, 2002, Abstr. 621 [2]), we have generated a combination plasmid, encoding the VEGF gene and the thymidine phosphorylase (TP, also known as platelet-derived endothelial growth factor (PD-ECGF) or gliostatin (GLS)) gene: phVEGF165-TP.MB. Upon transfection in COS-7 cells both gene products were expressed and functional as shown by Western blots, ELISAs and bioassays. Culture supernatants of COS-7 cells transfected with this plasmid were able to induce endothelial proliferation. In an in vitro angiogenesis assay with recombinant proteins, TP was able to increase VEGF-induced tube formation. The phVEGF165-TP.MB plasmid is therefore a promising candidate for in vivo angiogenesis studies.

摘要

为了改进目前使用编码血管内皮生长因子（VEGF）的质粒进行的血管生成基因治疗（Baumgartner等人，《循环》，1998年；97: 1114 - 23 [1]；Kusumanto等人，美国基因治疗学会第五届年会，波士顿，2002年，摘要621 [2]），我们构建了一种组合质粒，编码VEGF基因和胸苷磷酸化酶（TP，也称为血小板衍生内皮生长因子（PD - ECGF）或抑胶质瘤素（GLS））基因：phVEGF165 - TP.MB。如蛋白质免疫印迹、酶联免疫吸附测定和生物测定所示，转染到COS - 7细胞后，两种基因产物均有表达且具有功能。用该质粒转染的COS - 7细胞的培养上清液能够诱导内皮细胞增殖。在使用重组蛋白的体外血管生成测定中，TP能够增加VEGF诱导的管腔形成。因此，phVEGF165 - TP.MB质粒是体内血管生成研究的一个有前景的候选物。