Gene expression profile in corneal neovascularization identifiedby immunology related macroarray

[摘要] Purpose: To identify differentially expressed genes in cornealneovascularization using cDNA macroarray.Methods: Mechanical denudation of corneal and limbal epithelium wasperformed to induce corneal vascularization in mice. Corneas wereharvested 4 days after operation. Total RNA was isolated from bothnormal and vascularized corneas and used for the synthesis of cDNAprobes. 32P labeled exponential cDNA probes were hybridized tomouse cDNA immunology arrays. To validate the gene expression patternsrevealed by the cDNA expression array analysis, semiquantitative reversetranscription-polymerase chain reaction (RT-PCR) and statisticalanalysis were performed to compare the normal and vascularized cornealsamples.Results: Of 545 immunology related genes on the arrays, 6 genes wereupregulated and 1 gene was downregulated in the vascularized corneascompared with the normal corneas. Semi-quantitative RT-PCR was performedon the 6 genes selected in the arrays study, and showed thatcalreticulin (calregulin), apolipoprotein E, HSP84 (HSP90β), andpleiotrophin were upregulated while interferon regulatory factor-1 wasdownregulated in the vascularized corneas compared with the normalcorneas. These genes possess a number of biological functions includingmolecular chaperon, growth factor, and transcriptional factors.Conclusions: The differentially expressed genes newly identified inthe context of corneal neovascularization represent novel candidatefactors for further functional studies of the mechanisms of cornealneovascularization. Our data may provide new insight into the biologicalprocess of inflammation induced corneal neovascularization.

[学科分类] 生物化学/生物物理