Wilson S E, Walker J W, Chwang E L, He Y G
Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas.
Invest Ophthalmol Vis Sci. 1993 Jul;34(8):2544-61.
The purpose of this study was to determine whether messenger RNA coding for hepatocyte growth factor (HGF), HGF receptor (MET), keratinocyte growth factor (KGF), KGF receptor, and fibroblast growth factor (FGF) receptor-2 were produced in primary cultures of human corneal epithelial, stromal fibroblast, and endothelial cells, as well as ex vivo corneal epithelium, endothelial cells transfected with the SV40 large T antigen, and control embryonic lung fibroblasts. The effects of exogenous HGF and KGF, compared to epidermal growth factor, on the proliferation of first passage corneal cells were also examined.
Polymerase chain reaction was used to amplify complementary DNA for each modulator from each cell type. Hot blotting was used to demonstrate the specificity of amplification products. Proliferation of first passage corneal epithelial, stromal fibroblast, and endothelial cells in response to varying concentrations of HGF, KGF, and epidermal growth factor was measured.
Specific amplification products for messenger RNA coding for each modulator were detected in each corneal cell type, although very low levels of HGF and KGF messenger RNA appeared to be present in corneal epithelial cells relative to stromal fibroblasts and corneal endothelial cells. Amplification products that may have been derived from alternative transcripts were detected for several of the modulators. HGF and KGF stimulated proliferation in a dose-response manner in first passage corneal epithelial and endothelial cells, but not stromal fibroblast cells.
Human corneal epithelial, stromal fibroblasts, and endothelial cells produce messenger RNA coding for HGF and KGF, although low levels appear to be present in the epithelial cells. All three major cell types of the cornea produce messenger RNA coding for HGF receptor, KGF receptor, and FGF receptor-2. The proliferation of human corneal epithelial and endothelial cells, but not stromal fibroblasts, was stimulated by exogenous HGF and KGF. HGF and KGF likely have intracrine, autocrine, and/or paracrine functions in the cornea. Exogenous HGF and KGF may be useful in corneal preservation and for regulating corneal wound healing.
本研究旨在确定编码肝细胞生长因子（HGF）、HGF受体（MET）、角质形成细胞生长因子（KGF）、KGF受体和成纤维细胞生长因子（FGF）受体-2的信使核糖核酸是否在人角膜上皮细胞、基质成纤维细胞和内皮细胞的原代培养物中产生，以及在离体角膜上皮、转染了SV40大T抗原的内皮细胞和对照胚胎肺成纤维细胞中产生。还研究了与表皮生长因子相比，外源性HGF和KGF对第一代角膜细胞增殖的影响。
采用聚合酶链反应从每种细胞类型中扩增每种调节因子的互补DNA。热印迹法用于证明扩增产物的特异性。测量第一代角膜上皮细胞、基质成纤维细胞和内皮细胞对不同浓度的HGF、KGF和表皮生长因子的增殖反应。
在每种角膜细胞类型中均检测到编码每种调节因子的信使核糖核酸的特异性扩增产物，尽管相对于基质成纤维细胞和角膜内皮细胞，角膜上皮细胞中HGF和KGF信使核糖核酸的水平似乎非常低。检测到几种调节因子可能来自可变转录本的扩增产物。HGF和KGF以剂量反应方式刺激第一代角膜上皮细胞和内皮细胞的增殖，但不刺激基质成纤维细胞。
人角膜上皮细胞、基质成纤维细胞和内皮细胞产生编码HGF和KGF的信使核糖核酸，尽管上皮细胞中的水平似乎较低。角膜的所有三种主要细胞类型均产生编码HGF受体、KGF受体和FGF受体-2的信使核糖核酸。外源性HGF和KGF刺激人角膜上皮细胞和内皮细胞的增殖，但不刺激基质成纤维细胞。HGF和KGF可能在角膜中具有内分泌、自分泌和/或旁分泌功能。外源性HGF和KGF可能在角膜保存和调节角膜伤口愈合方面有用。
