[Analysis of IL-8 gene transcription in human bronchial epithelial cells by stable transfection of a reporter gene].
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.
Aoki
Y
Y
Department of Internal Medicine Saga Medical School, Japan.
D001980:Bronchi / Q000166:cytology*; D004847:Epithelial Cells / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection
D016209:Interleukin-8