Generation of an anti-angiogenic endothelial progenitor cell line via endostatin gene transfer

Ai,Jing; Sun,Jun‑Hui; Wan,Ting; Ma,Jian; Feng,Lei; Yao,Ke

doi:10.3892/mmr.2018.8623

Generation of an anti-angiogenic endothelial progenitor cell line via endostatin gene transfer

Authors:
- Jing Ai
- Jun‑Hui Sun
- Ting Wan
- Jian Ma
- Lei Feng
- Ke Yao
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Affiliations: Eye Centre, The Second Affiliated Hospital of The School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China, Key Laboratory of Combined Multi‑Organ Transplantation, Ministry of Public Health, The First Affiliated Hospital of The School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China
Published online on: February 21, 2018 https://doi.org/10.3892/mmr.2018.8623
Pages: 5814-5820
Copyright: © Ai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The viability of endothelial progenitor cells (EPCs) as a therapeutic treatment for neovascularization (NV) was subject to investigation in the present study. Furthermore, endostatin has previously been demonstrated to be an inhibitor of angiogenesis and a suppressant of vascular leakage. The aim of the present study was to generate transgenic EPCs with anti‑angiogenic effects for the treatment of ocular NV. EPCs were obtained from rat peripheral blood samples and then verified. A lentiviral‑endostatin‑green fluorescent protein recombinant construct was generated and used to infect EPCs. Transfected cells were then subjected to puromycin selection. Reverse transcription‑quantitative polymerase chain reaction and a western blot assay were then applied in order to determine both the endostatin mRNA and protein expression levels, respectively. In addition, vascular endothelial growth factor (VEGF) expression levels were also detected in order to observe the anti‑angiogenic effect of the endostatin‑transfected EPCs. Following puromycin (1 µg/ml) selection for 4 days, a stable endostatin‑transfected EPC line was generated. In this stable endostatin-transfected EPC line, the expression levels of endostatin increased; whereas the expression levels of VEGF decreased. The results of the present study revealed that EPCs can be genetically modified to overexpress endostatin, which may provide the cells with an anti‑angiogenic effect via increased expression of endostatin and decreased expression of VEGF. Thus, EPCs genetically modified to overexpress endostatin may serve as a potential therapeutic agent for ocular NV treatment.

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