| 摘要: |
| 目的 通过体外实验在角质细胞中过表达或沉默叉头框蛋白O1（FOXO1）基因，研究其对血管内皮生长因子A（VEGF-A）蛋白水平和转录活性的影响。方法 采用体外细胞培养和转染方法，分别用ON-TARGET plus SMART pool人FOXO1 siRNA和对照 siRNA（ON-TARGET plus Non-targeting Control Pool）转染人永生化牙龈角质细胞（human immortalized gingival keratinocyte，HIGK），用免疫荧光法检测HIGK中FOXO1和VEGF-A的免疫荧光强度，荧光酶素实验分别检测FOXO1过表达和FOXO1沉默时VEGF-A的活性。结果 与未转染组相比，使用FOXO1 siRNA转染的HIGK中VEGF-A的蛋白水平降低了57%（P<0.05）。荧光酶素实验分析结果显示，在HIGK中过表达FOXO1后VEGF-A的转录活性增加了2.1倍（P<0.05），而FOXO1沉默时VEGF-A的转录活性减少了20%~43%（P<0.05）。结论 角质细胞中FOXO1参与VEGF-A的表达调控。 |
| 关键词: 叉头框蛋白O1 血管内皮生长因子A 角质细胞 伤口愈合 |
| DOI：10.16781/j.0258-879x.2017.03.0318 |
| 投稿时间：2017-01-14修订日期：2017-02-27 |
| 基金项目:上海交通大学医学院附属新华医院基金（134525）. |
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| Forkhead box O1 gene in keratinocytes regulates expression of vascular endothelial growth factor-A |
| LU Yong-jian1,2,TANG Zhen1,WANG Da-lin1* |
(1. Department of Stomatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Stomatology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China
*Corresponding author) |
| Abstract: |
| Objective To explore the effect of forkhead box O1 (FOXO1) gene on the expression and transcription of vascular endothelial growth factor-A (VEGF-A) through overexpressing or silencing the FOXO1 gene in vitro. Methods The human immortalized gingival keratinocyte (HIGK) was cultured and transfected with ON-TARGET plus SMART pool for human FOXO1 siRNA or ON-TARGET plus Non-targeting Control Pool (control siRNA), the fluorescence intensity of VEGF-A was measured by the immunofluorescence staining in vitro. Luciferase assay was used to test the VEGF-A luciferase activity by FOXO1 overexpression or FOXO1 silence. Results The VEGF-A protein levels in HIGK transfected with FOXO1 siRNA significantly reduced by 57% compared with HIGK without transfection (P<0.05). The luciferase reporter results showed that overexpression of FOXO1 resulted in a 2.1-fold increase in VEGF-A transcriptional activity (P<0.05), and FOXO1 silencing produced a 20%-43% decrease in VEGF-A transcriptional activity (P<0.05). Conclusion FOXO1 in HIGKs can regulate VEGF-A protein expression. |
| Key words: forkhead box O1 vascular endothelial growth factor-A keratinocyte wound healing |
