[Expression of nuclear transcription factor NF-kappaB in human corneal fibroblasts].
OBJECTIVE
To explore the constitutive expression and activation of nuclear transcription factor NF-kappaB in corneal fibroblasts.


METHODS
2 or 3 passage corneal fibroblasts cultured in vitro were divided into two groups after synchronization. One group was cultured in DEME free of serum; the other in DEME with 10 microg/ml lipopolysaccharides for 6 hours. NF-kappaB was studied by flow cytometry (FCM) technique using immunofluorescent staining, and by gel shift assay (EMSA) using gamma-(32)P labeled NF-kappaB oligonucleotide probe.


RESULTS
According to FCM results, there were constitutively 9.4% NF-kappaB positive corneal fibroblasts, and 2.2 folds more (30.0%) after stimulated with lipopolysaccharides. Using EMSA method, we demonstrated the existence of NF-kappaB with DNA binding activity in the stimulated nuclei.


CONCLUSIONS
There is nuclear transcription factor NF-kappaB in human corneal fibroblasts. When activated, cells express more transcription factor which can enter nuclei, bind itself to the related DNAs and play a regulatory role of gene transcription.
Zhao
J
J
Department of Ophthalmology, The First Hospital of Beijing Medical University, Beijing 100034.
Wu
J
J
D002478:Cells, Cultured; D003319:Corneal Stroma / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D016328:NF-kappa B / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics
D008070:Lipopolysaccharides; D016328:NF-kappa B