Purchase this article with an account.
Tsan-Chi Chen, Shu-Wen Chang; Effect of Mitomycin C on IL-1R Expression, IL-1–Related Hepatocyte Growth Factor Secretion and Corneal Epithelial Cell Migration. Invest. Ophthalmol. Vis. Sci. 2010;51(3):1389-1396. doi: 10.1167/iovs.09-3494.
Download citation file:
© 2017 Association for Research in Vision and Ophthalmology.
To investigate how mitomycin C (MMC) modulates hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) secretions in human corneal fibroblasts and regulates human corneal epithelial (HCE) cell migration.
Primary human corneal fibroblasts were treated with MMC (0.05, 0.1, or 0.2 mg/mL for 5 minutes) and were cultivated with or without interleukin (IL)-1β. Transcript and secretion of HGF and KGF were determined by quantitative real-time RT-PCR and Western blot analysis, respectively. The effect of MMC-treated fibroblasts on HCE cell migration was evaluated using a transwell migration assay. The influence of MMC on HGF expression/secretion and HCE cell migration was further confirmed by RNA interference. The number of IL-1 receptors (IL-1R) on the fibroblast surface was analyzed by flow cytometry.
MMC alone did not affect endogenous HGF expression, whereas IL-1β alone significantly upregulated HGF transcripts and secretion. By modifying IL-1R numbers, MMC further upregulated IL-1β–related HGF expression at a concentration of 0.05 mg/mL but to a lesser extent at 0.1 and 0.2 mg/mL. KGF transcripts and intracellular expression were suppressed by MMC dose dependently in the presence or absence of IL-1β, whereas KGF secretion was not affected. Conditioned medium from MMC-treated fibroblasts exerted a similar concentration-dependent effect on HCE cell migration, enhancing migration most significantly at 0.05 mg/mL MMC in the presence of IL-1β. The MMC dose-dependent modulation of HCE cell migration was abolished in HGF-silenced fibroblasts.
MMC differentially modulated IL-1R expression at various concentrations and regulated HGF and KGF differently. MMC alone did not alter HGF expression. In the presence of IL-1β, MMC-treated corneal fibroblasts modified HCE cell migration through IL-1β–induced HGF secretion.
This PDF is available to Subscribers Only