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腺病毒介导的外源性Tum5基因的表达对脐静脉内皮细胞的抑制作用

发表时间:2016-09-15  浏览量:1525  下载量:341
全部作者: 贾育蓉,杨伟,张红,张琰,孙靖
作者单位: 天津医科大学眼科医院,天津医科大学眼科研究所,天津医科大学眼视光学院
摘 要: 目的:研究腺病毒介导Tum5重组基因对生理状态下人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖、迁移及管腔成形的影响。方法:构建表达绿色荧光蛋白的空载体腺病毒(rAd-GFP)和携带Tum5基因的重组腺病毒(rAd-Tum5)。分别以1×1010 cells/mL的病毒颗粒感染HUVEC,将HUVEC分为正常对照组、空载体对照组(rAd-GFP组)和Tum5基因组(rAd-Tum5组)。采用细胞计数试剂盒(CCK-8)、侵袭小室(Transwell)实验及基质胶(Matrigel)实验分析Tum5重组基因对HUVEC增殖、迁移和管腔成形的影响。收集转染后24,48,72 h的细胞上清液,采用人VEGF ELISA试剂盒,检测上清液中VEGF含量的变化。结果:CCK-8、Transwell及Matrigel实验结果显示,正常对照组、rAd-GFP组和rAd-Tum5组3组间细胞的增殖数量、迁移数量及管腔成形的数量比较,差异均有统计学意义(F=78.914,8.524,226.498,P<0.05)。细胞增殖数量、迁移数量及管腔成形数量组间两两比较结果显示,正常对照组与rAd-GFP组之间无明显差异(P>0.05),而rAd-Tum5组较正常对照组、rAd-GFP组显著减少,差异有统计学意义(P<0.05)。ELISA结果显示,rAd-Tum5组较正常对照组、rAd-GFP组上清液中VEGF的含量显著减少(P<0.05),而正常对照组、rAd-GFP组中VEGF的含量无显著性差异(P>0.05)。结论:腺病毒介导Tum5重组基因可抑制生理状态下HUVEC的增殖、迁移及管腔成形,这可能与Tum5抑制HUVEC分泌VEGF有关。
关 键 词: 眼科学;Tum5;内皮细胞;细胞增殖;细胞迁移;管腔成形
Title: Inhibitory effects of adenovirus-mediated overexpression of exogenous Tum5 gene on human umbilical endothelial cells
Author: JIA Yurong, YANG Wei, ZHANG Hong, ZHANG Yan, SUN Jing
Organization: Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, College of Optometry and Ophthalmology, Tianjin Medical University
Abstract: Objective: To study the effects of adenovirus-mediated overexpression of exogenous Tum5 gene on the proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVEC). Methods: The empty adenoviral vector expressing green fluorescent protein (rAd-GFP) and the adenoviral vector expressing the recombinant Tum5 gene (rAd-Tum5) were constructed and packaged into the viral particles. HUVEC were divided into normal group, empty expression vector (rAd-GFP) group, and rAd-Tum5 group. The rAd-GFP and rAd-Tum5 groups of cells were infected with the corresponding viral particles (1×1010cells/mL), and the normal group was left untreated. The CCK-8 test was used for examining the cell proliferation, the Transwell test for the cell migration, and the Matrigel test for the tube formation of the cells. The cell supernatants at 24, 48, and 72 h after viral infection were collected and examined for the protein levels of VEGF by a Human VEGF ELISA kit. Results: The results of CCK-8, Transwell and Matrigel tests showed that there were statistical differences between all the experimental groups in the proliferation, migration and tubing of the HUVEC (F=78.914, 8.524, 226.498, all P<0.05 ). There were no statistical differences between normal group and rAd-GFP group (all P>0.05). However, these parameters in the rAd-Tum5 group were significantly lower than the normal and the rAd-GFP groups (all P<0.05). ELISA results showed that the VEGF protein levels in the supernatant of the rAd-Tum5 group were significantly reduced compared to those in the normal group and rAd-GFP group (all P<0.05). There were no statistical differences between the normal group and the rAd-GFP group in the protein levels of VEGF (P>0.05). Conclusion: The adenoviral vector-mediated overexpression of exogenous Tum5 gene can inhibit the proliferation, migration, and tube formation of HUVEC, which may be due to the Tum5-mediated inhibition of VEGF protein levels.
Key words: ophthalmology; Tum5; endothelial cells; cell proliferation; cell migration; tube formation
发表期数: 2016年9月第17期
引用格式: 贾育蓉,杨伟,张红,等. 腺病毒介导的外源性Tum5基因的表达对脐静脉内皮细胞的抑制作用[J]. 中国科技论文在线精品论文,2016,9(17):1778-1784.
 
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