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慢病毒介导的内皮抑素对内皮祖细胞的影响。

Effects of lentivirus-mediated endostatin on endothelial progenitor cells.

作者信息

Ai Jing, Sun Jun-Hui, Ma Jian, Yao Ke

机构信息

Eye Center, 2nd Affiliated Hospital of School of Medicine, Zhejiang University, Hangzhou 310009, China.

Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China.

出版信息

Oncotarget. 2017 Oct 10;8(55):94431-94439. doi: 10.18632/oncotarget.21770. eCollection 2017 Nov 7.

DOI:10.18632/oncotarget.21770
PMID:29212239
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5706885/
Abstract

Endothelial progenitor cells (EPCs) are candidates for gene therapies against retinal neovascularization (NV). The aim of present study was to investigate the effects of endostatin transfection on EPC function. In the present study, the EPCs were infected with lentivirus overexpressing endostatin. The transfection effects of endostatin overexpression on the proliferation, migratory, differentiation, apoptosis and the cell cycle of this cell line were determined. The real-time quantitative polymerase chain reaction (RT-qPCR) and western blot assays showed high expression levels of endostatin. A cell counting kit-8 assay showed that endostatin overexpression inhibited EPC proliferation. The transwell assay indicated that endostatin overexpression could suppress EPC migration. Furthermore, endostatin overexpression enhanced apoptosis (as showed by AnnexinV-FITC/propidiumiodide staining analysis), induced differentiation and blocked the cell cycle. As compared with negative control group, EPC viability significantly decreased in gene transfection group. In conclusion, present study determined the feasibility of lentivirus-mediated endostatin gene transfer, and indirectly proved the effect of endostatin secretion on EPCs. Also our study provided a new opportunity for the potential application of gene therapy in retinal NV.

摘要

内皮祖细胞（EPCs）是针对视网膜新生血管形成（NV）进行基因治疗的候选细胞。本研究的目的是探讨内皮抑素转染对EPC功能的影响。在本研究中，EPCs被过表达内皮抑素的慢病毒感染。测定了内皮抑素过表达对该细胞系增殖、迁移、分化、凋亡及细胞周期的转染效果。实时定量聚合酶链反应（RT-qPCR）和蛋白质印迹分析显示内皮抑素表达水平较高。细胞计数试剂盒-8检测显示内皮抑素过表达抑制EPC增殖。Transwell检测表明内皮抑素过表达可抑制EPC迁移。此外，内皮抑素过表达增强了凋亡（通过膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色分析显示），诱导了分化并阻断了细胞周期。与阴性对照组相比，基因转染组EPC活力显著降低。总之，本研究确定了慢病毒介导的内皮抑素基因转移的可行性，并间接证明了内皮抑素分泌对EPCs的作用。我们的研究也为基因治疗在视网膜NV中的潜在应用提供了新机会。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/9ead668c1399/oncotarget-08-94431-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/34a60a53fff0/oncotarget-08-94431-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/9e25cf34e7e6/oncotarget-08-94431-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/813c9214f061/oncotarget-08-94431-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/79140bc1e5f0/oncotarget-08-94431-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/8b256983f605/oncotarget-08-94431-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/9ead668c1399/oncotarget-08-94431-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/34a60a53fff0/oncotarget-08-94431-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/9e25cf34e7e6/oncotarget-08-94431-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/813c9214f061/oncotarget-08-94431-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/79140bc1e5f0/oncotarget-08-94431-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/8b256983f605/oncotarget-08-94431-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6bf/5706885/9ead668c1399/oncotarget-08-94431-g006.jpg

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