Hypertonic stress induces c-fos but not c-jun expression in the human embryonal EUE epithelial cell line.
Recent evidence has indicated a role for the two early response genes c-fos and c-jun in transcriptional regulation of genes acting in osmoregulation. On this basis we investigated their expression in response to hypertonic stress in the human embryonal EUE epithelial cell line. EUE cells have proven to be a useful tool for studying long-term in vitro adaptation to hypertonic stress. After culturing EUE cells in hypertonic medium a marked c-fos induction was observed, both at the mRNA and the protein level. Northern analysis of fos-mRNA showed a peak expression at 4 h, followed by a progressive decline till complete extinction at 8 h. Immunofluorescence analysis of FOS protein evidenced a similar, although slightly delayed kinetics of expression. Conversely, neither c-jun nor c-myc up-regulation could be detected. The treatment of EUE cells with cycloheximide led to superinduction of c-fos expression, (with high levels up to 12 h), and to a c-jun expression that was just detectable. Hypertonic stimulation of the transformed cell lines A549, MCF7 and JR induced both c-fos and c-jun only in JR cells. Hypertonic shock was also effective in inducing c-fos expression in fetal human diploid fibroblasts, although the response was earlier and more transient than in EUE cells. These findings indicate that c-fos is a primary response gene in hypertonic stress-activated cells, although the pattern and kinetics of its induction may differ according to the type of cell.
Rossi
D
D
Department of Biology and Genetics, University of Milan, Italy.
Fuhrman Conti
A M
AM
De Grada
L
L
Larizza
L
L
D015152:Blotting, Northern; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D015342:DNA Probes; D004848:Epithelium / Q000378:metabolism; D005786:Gene Expression Regulation; D016762:Genes, fos; D016758:Genes, jun; D006801:Humans; D006982:Hypertonic Solutions / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D013997:Time Factors
D015342:DNA Probes; D006982:Hypertonic Solutions; D003513:Cycloheximide