CTCF 在表皮生长因子诱导的永生化人角膜上皮细胞迁移中的作用。
Role of CTCF in EGF-induced migration of immortalized human corneal epithelial cells.
机构信息
Department of Medicine, Jules Stein Eye Institute, David Geffen School of Medicine, University of California Los Angeles, CA 90502, USA.
出版信息
Invest Ophthalmol Vis Sci. 2012 Feb 23;53(2):946-51. doi: 10.1167/iovs.11-8747. Print 2012 Feb.
PURPOSE
EGF-induced activation of the epigenetic CCCTC binding factor (CTCF) plays an important role in corneal epithelial cell proliferation by suppressing the Pax6 gene. The present study focused on further understanding the role of CTCF in mediating EGF-induced migration of immortalized human corneal epithelial cells.
METHODS
CTCF activities in human corneal epithelial cells immortalized by telomerase (HTCE cells) and SV-40 (HCE cells) transformation were suppressed and enhanced by CTCF mRNA knockdown and by overexpressing CTCF cDNA, respectively. EGF-induced cell migration was evaluated by linear scratch wound healing, a cell migration assay, and live cell motility GFP-tracking with a fluorescence microscope. Immunochemical analysis was performed for detecting focal adhesion changes in EGF-induced and CTCF activity-altered cells.
RESULTS
EGF-induced wound closure and cell migration rates of human corneal epithelial cells were significantly suppressed and enhanced by CTCF mRNA knockdown and by overexpression of CTCF, respectively. CTCF mRNA knockdown also markedly suppressed cell motility, determined by using a live-cell-tracking system in GFP-tag-expressed HTCE cells. Finally, alterations of EGF-stimulated focal adhesion were observed in CTCF knockdown HTCE cells by immunostaining of F-actin and vinculin in cytoskeleton reorganization.
CONCLUSIONS
CTCF, an epigenetic regulator and transcription factor, involves EGF-induced increases in cell motility and migration. CTCF plays an essential role in growth factor-regulated human corneal epithelial cell wound healing.
目的
表皮生长因子（EGF）诱导的表观遗传 CCCTC 结合因子（CTCF）的激活通过抑制 Pax6 基因在角膜上皮细胞增殖中发挥重要作用。本研究旨在进一步了解 CTCF 在介导永生化人角膜上皮细胞中 EGF 诱导的迁移中的作用。
方法
通过 CTCF mRNA 敲低和过表达 CTCF cDNA 分别抑制和增强端粒酶（HTCE 细胞）和 SV-40（HCE 细胞）转化的人角膜上皮细胞中的 CTCF 活性。通过线性划痕愈合、细胞迁移测定和荧光显微镜下 GFP 标记的活细胞运动追踪评估 EGF 诱导的细胞迁移。进行免疫化学分析以检测 EGF 诱导和 CTCF 活性改变细胞中的焦点粘连变化。
结果
EGF 诱导的人角膜上皮细胞的伤口闭合和细胞迁移率分别通过 CTCF mRNA 敲低和 CTCF 的过表达显著抑制和增强。CTCF mRNA 敲低也显著抑制了 GFP 标记的 HTCE 细胞中活细胞追踪系统测定的细胞运动。最后，通过细胞骨架重排中的 F-肌动蛋白和纽蛋白的免疫染色观察到 CTCF 敲低 HTCE 细胞中 EGF 刺激的焦点粘连的改变。
结论
作为一种表观遗传调节剂和转录因子，CTCF 参与 EGF 诱导的细胞迁移和迁移增加。CTCF 在生长因子调节的人角膜上皮细胞伤口愈合中起重要作用。
Zotero 插件