Wilson S E, He Y G, Weng J, Zieske J D, Jester J V, Schultz G S
Department of Ophthalmology, University of Texas Southwestern Medical Center at Dallas 75235, USA.
Exp Eye Res. 1994 Dec;59(6):665-78. doi: 10.1006/exer.1994.1152.
We sought to determine the effects of exogenous epidermal growth factor (EGF), heparin-binding EGF (HB-EGF), transforming growth factor alpha (TGF-alpha), single-chain precursor hepatocyte growth factor (SC-HGF), double-chain mature HGF (DC-HGF), and keratinocyte growth factor (KGF) on proliferation, motility, and differentiation of first passage cultures of human corneal epithelial cells in serum-free chemically defined medium. The effect of EGF, HB-EGF, TGF-alpha, SC-HGF, DC-HGF, KGF or combinations of the growth factors on proliferation was measured by counting cells present after 3 weeks of culture and by immunostaining for the cell-cycle-specific nuclear proliferation antigen Ki-67. The effect of the factors on epithelial cell motility was assessed by morphometric analysis of photographs of cells migrating from confluent islands of cells. The effect of growth factors on differentiation of epithelial cells were determined by immunostaining epithelial cell islands for the keratin K3 and by Western blotting for keratin K3. EGF, alone or in combination with KGF and SC-HGF, significantly stimulated motility of epithelial cells at the periphery of confluent islands of cells and induced an elongated cell morphology. TGF-alpha, HB-EGF and DC-HGF produced motility effects similar to EGF. There was diminished proliferation of the migrating cells in response to EGF, HB-EGF, TGF-alpha or DC-HGF, while non-migrating epithelial cells in the center of confluent islands continued to proliferate in response to the growth factors. EGF, HB-EGF, TGF alpha or DC-HGF inhibited expression of the differentiation-related marker keratin K3 in epithelial cells, both at the edge and at the center of the islands. KGF stimulated proliferation of corneal epithelial cells at low density and in confluent islands of cells. KGF did not affect expression of keratin K3 or migration of epithelial cells. SC-HGF had no effect on corneal epithelial cells. These results indicate that the effects of EGF, HB-EGF, TGF-alpha and DC-HGF on corneal epithelial cell proliferation, motility and differentiation vary from those of KGF and SC-HGF. EGF, HB-EGF, TGF-alpha and DC-HGF induced changes in epithelial cell morphology and motility in cells plated at low cell density or in cells located at the edge of a confluent island. Thus, these effects appear to be dependent on the extent of cell-cell contact. The inhibitory effect of EGF, HB-EGF, TGF-alpha or DC-HGF on corneal epithelial cell differentiation, however, is independent of cell density.(ABSTRACT TRUNCATED AT 400 WORDS)
我们试图确定外源性表皮生长因子（EGF）、肝素结合表皮生长因子（HB-EGF）、转化生长因子α（TGF-α）、单链前体肝细胞生长因子（SC-HGF）、双链成熟肝细胞生长因子（DC-HGF）和角质形成细胞生长因子（KGF）对无血清化学限定培养基中第一代人角膜上皮细胞培养物的增殖、迁移和分化的影响。通过计数培养3周后存在的细胞以及对细胞周期特异性核增殖抗原Ki-67进行免疫染色，来测定EGF、HB-EGF、TGF-α、SC-HGF、DC-HGF、KGF或这些生长因子组合对增殖的影响。通过对从汇合细胞岛迁移的细胞照片进行形态计量分析，评估这些因子对上皮细胞迁移的影响。通过对角质形成细胞岛进行角蛋白K3免疫染色以及对K3进行蛋白质印迹分析，来确定生长因子对上皮细胞分化的影响。EGF单独或与KGF和SC-HGF联合使用时，能显著刺激汇合细胞岛周边上皮细胞的迁移，并诱导细胞形态拉长。TGF-α、HB-EGF和DC-HGF产生与EGF相似的迁移效应。对EGF、HB-EGF、TGF-α或DC-HGF有反应的迁移细胞增殖减少，而汇合细胞岛中心的非迁移上皮细胞对生长因子仍有增殖反应。EGF、HB-EGF、TGF-α或DC-HGF在细胞岛边缘和中心均抑制上皮细胞中与分化相关的标志物角蛋白K3的表达。KGF在低密度和汇合细胞岛中刺激角膜上皮细胞增殖。KGF不影响角蛋白K3的表达或上皮细胞的迁移。SC-HGF对角膜上皮细胞无影响。这些结果表明，EGF、HB-EGF、TGF-α和DC-HGF对角膜上皮细胞增殖、迁移和分化的影响与KGF和SC-HGF不同。EGF、HB-EGF、TGF-α和DC-HGF在低密度接种的细胞或位于汇合细胞岛边缘的细胞中诱导上皮细胞形态和迁移的变化。因此，这些效应似乎取决于细胞间接触的程度。然而，EGF、HB-EGF、TGF-α或DC-HGF对角膜上皮细胞分化的抑制作用与细胞密度无关。（摘要截断于400字）
