Effects of lentivirus-mediated endostatin on endothelial progenitor cells.
Effects of lentivirus-mediated endostatin on endothelial progenitor cells.
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慢病毒介导的内皮抑素对内皮祖细胞的影响
DOI：
10.18632/oncotarget.21770
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发表时间：
2017-11-07
期刊：
影响因子：
--
通讯作者：
Yao K
中科院分区：
文献类型：
--
作者：
Ai J;Sun JH;Ma J;Yao K
内皮祖细胞（EPCs）是视网膜新生血管（NV）基因治疗的候选细胞。本研究旨在探讨内皮抑素基因转染对内皮祖细胞功能的影响。在本研究中，EPCs被过表达内皮抑素的慢病毒感染。检测内皮抑素过表达对该细胞系增殖、迁移、分化、凋亡及细胞周期的影响。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的高表达水平。细胞计数kit-8法显示内皮抑素过表达抑制EPC增殖。transwell法显示内皮抑素过表达可抑制EPC迁移。此外，内皮抑素过表达增强凋亡（如AnnexinV-FITC/碘化丙啶染色分析所示），诱导分化并阻断细胞周期。与阴性对照组相比，基因转染组EPC活性明显降低。本研究证实了慢病毒介导的内皮抑素基因转移的可行性，并间接证实了内皮抑素分泌对内皮祖细胞的影响。本研究为基因治疗在视网膜NV中的应用提供了新的契机。
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.
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