Sex hormone modulation of cell growth and apoptosis of the human monocytic/macrophage cell line.
Sex hormones seem to modulate the immune/inflammatory responses by different mechanisms in female and male rheumatoid arthritis patients. The effects of 17beta-oestradiol and of testosterone were tested on the cultured human monocytic/macrophage cell line (THP-1) activated with IFN-gamma in order to investigate their role in cell proliferation and apoptosis. Activated human THP-1 cells were cultured in the presence of 17beta-oestradiol and testosterone (final concentration, 10 nM). The evaluation of markers of cell proliferation included the NF-kappaB DNA-binding assay, the NF-kappaB inhibition complex, the proliferating cell nuclear antigen expression and the methyl-tetrazolium salt test. Apoptosis was detected by the annexin V-propidium assay and by the cleaved poly-ADP ribose polymerase expression. Specific methods included flow analysis cytometry scatter analysis, immunocytochemistry and western blot analysis. Cell growth inhibition and increased apoptosis were observed in testosterone-treated THP-1 cells. Increased poly-ADP ribose polymerase-cleaved expression and decreased proliferating cell nuclear antigen expression, as well as an increase of IkappaB-alpha and a decrease of the IkappaB-alpha phosphorylated form (ser 32), were found in testosterone-treated THP-1 cells. However, the NF-kappaB DNA binding was found increased in 17beta-oestradiol-treated THP-1 cells. The treatment with staurosporine (enhancer of apoptosis) induced decreased NF-kappaB DNA binding in all conditions, but particularly in testosterone-treated THP-1 cells. Treatment of THP-1 by sex hormones was found to influence cell proliferation and apoptosis. Androgens were found to increase the apoptosis, and oestrogens showed a protective trend on cell death--both acting as modulators of the NF-kappaB complex.
Cutolo
Maurizio
M
Research Laboratory and Division of Rheumatology, Department of Internal Medicine, University of Genova, Italy. mcutolo@unige.it
Capellino
Silvia
S
Montagna
Paola
P
Ghiorzo
Paola
P
Sulli
Alberto
A
Villaggio
Barbara
B
D017209:Apoptosis / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D004958:Estradiol / Q000494:pharmacology*; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects*; D009000:Monocytes / Q000187:drug effects*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D019311:Staurosporine / Q000494:pharmacology; D013739:Testosterone / Q000494:pharmacology*
D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D018809:Proliferating Cell Nuclear Antigen; D000072000:NF-KappaB Inhibitor alpha; D013739:Testosterone; D004958:Estradiol; D007371:Interferon-gamma; D019311:Staurosporine