Abstract
The purpose of this study was to characterize the pattern of cytokine gene expression by human corneal epithelial cells (HCEC) in response to interleukin-1 (IL-1). Primary cultured HCEC (P-HCEC) or SV40 transformed HCEC (SV40-HCEC) were treated for 6 hr with serum-free growth-media alone or with recombinant human IL-1β or IL-1α (10 ng ml−1). 33P labeled cDNA was generated from total RNA, then hybridized to a human cytokine expression array. An autoradiograph was generated for each experimental condition and results analysed semi-quantitatively. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect mRNA for IL-8, growth related oncogene-β (GRO-β), intercellular adhesion molecule (ICAM)-1 and Ephrin A5. P-HCEC and SV40-HCEC demonstrated comparable cytokine profiles. For P-HCEC (n=2) the expression of 35 genes was upregulated or only detectable following IL-1β treatment whereas the expression of nine genes was downregulated or undetectable after IL-1β treatment. In SV40-HCEC (n=3), the expression of 48 genes was upregulated or only detectable following IL-1β treatment and the expression of 10 genes was downregulated or undetectable after IL-1β treatment. Some genes that demonstrated increased expression included cadherin-5, ICAM-1, GRO-α, GRO-β, GRO-γ, Activin A (bA subunit), tumor necrosis factor-α, IL-6, and IL-8. Genes that showed decreased expression included the chemokine receptor—CXCR-4, ciliary neurotrophic factor (CNTF), c-kit ligand, Ephrin A5, G-protein coupled receptor RDC-1 and FGF family FGFR2. Bayesian analysis of the SV40-HCEC data (n=3) revealed the expression of 15 genes that were significantly (p
Original language | English |
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Pages (from-to) | 175-183 |
Journal | Experimental Eye Research |
Volume | 80 |
Issue number | 2 |
Early online date | 27 Oct 2004 |
DOIs | |
Publication status | Published - Feb 2005 |
Keywords
- cornea
- epithelial cells
- gene expression
- interleukin-1
- cytokines
- inflammation
- ocular surface
- cell culture