Hydrogen peroxide-induced expression of the proto-oncogenes, c-jun, c-fos and c-myc in rabbit lens epithelial cells.

Abstract

The involvement of H2O2 in cataract development has been established in both human patients and animal models. At the molecular level H2O2 has been observed to cause damage to DNA, protein and lipid. To explore the oxidative stress response of the lens system at the gene expression level, we have examined the effects of H2O2 on the mRNA change of the proto-oncogenes, c-jun, c-fos and c-myc in a rabbit lens cell line, N/N1003A. H2O2 treatment of the rabbit lens epithelial cells for 60 min induces quick up-regulation of both c-jun and c-fos mRNAs. The maximal induction is 38 fold for c-jun at 150 microM and 72 fold for c-fos at 250 microM H2O2. Treatment of N/N1003A cells with 50-250 microM H2O2 for 60 min leads to a 2-5 fold increase of the c-myc mRNA level. H2O2 also induces an up-regulation in transactivity of the activating protein-1 (AP-1) as shown with a reporter gene driven by a prolactin gene promoter with 4 copies of AP-1 binding sites inserted in the upstream of the promoter. Maximal induction occurs with 150 microM H2O2. In the same system, the antioxidants, N-acetyl-cysteine (NAC) and pyrrolidine dithiocarbamate (PDTC) at concentrations shown to up-regulate the mRNAs of both c-jun and c-fos, also enhance the transactivity of AP-1. NAC and PDTC have different effects in modulating the induction of AP-1 activity by H2O2 and TPA. These results reveal that oxidative stress regulates expression of various regulatory genes in lens systems, which likely affects cell proliferation, differentiation and viability and thus affect normal lens functions.