Steroid-regulated growth of DDT1MF-2 cells is profoundly influenced by culture conditions |
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Authors: | Dolores J Lamb Madhumita Ray |
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Institution: | (1) Scott Department of Urology, Baylor College of Medicine, One Baylor Plaza, 77030 Houston, Texas;(2) The Department of Cell Biology, Baylor College of Medicine, One Baylor Plaza, 77030 Houston, Texas |
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Abstract: | Summary DDT1MF-2 cells provide an ideal model for studying tumor-growth-stimulation by steroids. These cells progress to a rapidly proliferating,
androgen-independent state after prolonged culture without androgen. After brief culture in different lots of fetal bovine
serum (FBS), some lots induced a permanent state of hormone-independence in cells that had been androgen-responsive. To test
the hypothesis that factors influenced androgen-responsive growth even after removal of serum, hormone-responsive DDT1MF-2 cells (7000 cells/well) were plated in medium Dulbecco’s Modified Eagle Medium/F-12 Nutrition Mixture (1:1)/1% ITS with
(a) 0.1% FBS, (b) 0.1% NuSerum (c) 0.1% Hyclone, or (d) MCDB-110/0.1% ITS with 5 ng/ml bFGF. On Days 2–8, medium was replaced
with D-MEM/F12/ITS with 10 nM testosterone (T), 10 nM triamcinolone acetonide (TA), or ethanol (control) and the cells counted. While testosterone induced a 1.4-fold increase
in cell growth after exposure to FBS or NuSerum, maximal testosterone effect (3-6-fold increase) was observed after Hyclone.
Hydroxyflutamide antagonized the fivefold increase in growth observed with testosterone, with a slight decrease of growth
with cAMP for cells plated in Hyclone. Androgen-independent cells were unaffected by testosterone, hydroxyflutamide, or 8Br-cAMP
medium (a)]. Maximal inhibition by triamcinolone acetonide (0.25 of control) was observed with medium (d). The effect of
testosterone and triamcinolone acetonide on secretion of mitogenic activity into conditioned medium was also evaluated. Although
conditioned media from control and testosterone-treated cells were mitogenic in a dose-dependent manner, the media from cells
treated with triamcinolone acetonide and testosterone + TA conditioned medium was not mitogenic—but, of note, it was not growth
inhibitory. |
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Keywords: | steroid growth regulation male reproduction tumor |
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