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Analysis of IL-8 gene transcription in human bronchial epithelial cells by stable transfection of a reporter gene

Aoki, Y.

Nihon Kyobu Shikkan Gakkai Zasshi 35(6): 602-608

1997

ISSN/ISBN: 0301-1542
PMID: 9294292
Document Number: 483241
Transcriptional activation of the expression of the gene for interleukin (IL)-8) in airway epithelial cells was analyzed by stable transfection of human bronchial epithelial cells (16 HBE) with a reporter plasmid pIL-8/Luc, which the expression of luciferase reporter gene is driven by human IL-8 promoter. As compared with the resting condition, the luciferase activity was 3.9 times higher after stimulation with phorbol-myristate-acetate (20 ng/ml), 1.6 times higher after stimulation with tumor necrosis factor-alpha (100 U/ml), and 2.7 times higher after stimulation with interleukin-1 beta (100 U/ml). Dexamethasone inhibited the effects of these stimulants by 10 to 50 percent. These results closely correspond to those of IL-8 mRNA analyses with Northern blotting and IL-8 protein analyses done by enzyme-linked immunosorbent assay. The transgenic cell line IL-8 Luc/16HBE can also be used in screening for drug effects, and may be useful for quantifying IL-8 inducibility in clinical samples obtained from the lungs.

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