Paracrine role of keratinocyte growth factor in rabbit corneal epithelial cell growth.
Keratinocyte growth factor (KGF), a member of the fibroblast growth factor (FGF) family, is expressed in stromal fibroblasts and acts specifically on cells of epithelial origin as a paracrine mediator. In this study, we examined KGF and KGF receptor (KGF-R) mRNA expression in rabbit corneal epithelial cells and keratocytes, using a reverse transcription polymerase chain reaction (RT-PCR) method. We also examined the effects of KGF on the proliferation of rabbit corneal cells in vitro. mRNA from cultured rabbit skin fibroblasts, conjunctival fibroblasts, corneal epithelial cells and keratocytes was first reverse-transcribed, and then amplified by PCR using primers specific for KGF and KGF-R. KGF mRNA was expressed in skin fibroblasts, conjunctival fibroblasts and keratocytes, but not in corneal epithelial cells. In contrast, KGF receptor mRNA was expressed in corneal epithelial cells, but not in skin fibroblasts, conjunctival fibroblasts or keratocytes. The results of Southern hybridization experiments of PCR products also showed that KGF mRNA was expressed in skin fibroblasts, conjunctival fibroblasts and keratocytes, whereas KGF receptor mRNA was expressed in corneal epithelial cells. Nucleotide sequencing of the PCR products revealed that their sequences were identical to the expected nucleotide sequences. Using the methylene blue method, we found that KGF enhanced the growth of cultured rabbit corneal epithelial cells, but not keratocytes, at a concentration of 0.01 to 100 ng ml-1. Furthermore, KGF also augmented 3H-thymidine incorporation into the corneal epithelial cells. These results suggest that KGF is a paracrine mediator of the proliferation of corneal epithelial cells as well as of keratinocytes in the skin.
Sotozono
C
C
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Japan.
Kinoshita
S
S
Kita
M
M
Imanishi
J
J
D000818:Animals; D002478:Cells, Cultured; D003315:Cornea / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D004848:Epithelium / Q000378:metabolism; D051526:Fibroblast Growth Factor 10; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors; D005347:Fibroblasts / Q000378:metabolism; D006133:Growth Substances / Q000096:biosynthesis* / Q000494:pharmacology; D015603:Keratinocytes / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017468:Receptors, Fibroblast Growth Factor; D017978:Receptors, Growth Factor / Q000096:biosynthesis
D051526:Fibroblast Growth Factor 10; D006133:Growth Substances; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; D017978:Receptors, Growth Factor; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors; D004247:DNA; D051497:Receptor, Fibroblast Growth Factor, Type 2; C065104:keratinocyte growth factor receptor