Up-regulation of semaphorin 3A in human corneal fibroblasts by epidermal growth factor released from cocultured human corneal epithelial cells.
Semaphorins are a family of glycoproteins that play an important role in repulsive axon guidance during embryogenesis. We have now investigated the effect of corneal epithelial cells on the expression of Sema3A in corneal fibroblasts with the use of a coculture system in which the two cell types are separated by a collagen vitrigel membrane. Reverse transcription-polymerase chain reaction and immunoblot analyses revealed that the presence of immortalized human corneal epithelial (HCE) cells increased the expression of Sema3A in human corneal fibroblasts at both the mRNA and protein levels. This effect of HCE cells was mimicked by recombinant human epidermal growth factor (EGF) in a concentration- and time-dependent manner. An inhibitor of the tyrosine kinase activity of the EGF receptor, PD153035, blocked the EGF-induced up-regulation of both Sema3A mRNA and protein in corneal fibroblasts. Depletion of EGF in HCE cells by RNA interference largely abolished the effect of these cells on Sema3A expression in corneal fibroblasts. These findings indicate that EGF released from corneal epithelial cells up-regulates the expression of Sema3A in corneal fibroblasts. This effect of EGF may play an important role in maintenance of corneal structure and repair of corneal damage.
Ko
Ji-Ae
JA
Department of Ophthalmology, Yamaguchi University Graduate School of Medicine, Ube City, Yamaguchi, Japan. jiae0831@yamaguchi-u.ac.jp
Morishige
Naoyuki
N
Yanai
Ryoji
R
Nishida
Teruo
T
D002460:Cell Line; D018920:Coculture Techniques; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019573:Epithelium, Corneal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D034622:RNA Interference; D011994:Recombinant Proteins / Q000494:pharmacology; D040121:Semaphorin-3A / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation
D011994:Recombinant Proteins; C108639:SEMA3A protein, human; D040121:Semaphorin-3A; D004815:Epidermal Growth Factor