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Differential effects of Resveratrol on androgen-responsive LNCaP human prostate cancer cells in vitro and in vivo. P-4

Materials and methods

Chemicals

Resveratrol, 17β-estradiol, dihydrotestosterone and dimethylsulfoxide were from Sigma Chemical Co. (St Louis, MO). The synthetic androgen R1881 was from NEN Life Science Products (Boston, MA).

Cells and cell culture

LNCaP human prostate cancer cells were obtained from the American Type Culture Collection (Manassas, VA) and maintained in Media A [RPMI 1640 medium with phenol red (Invitrogen, Carlsbad, CA), 2 mM L-glutamine (Sigma, St. Louis, MO), 100 U/ml penicillin and 100 μg/ml streptomycin (BioSource International, Camarillo, CA) with 10% fetal bovine serum (FBS) (Invitrogen)]. Cells were incubated in the presence of 5% CO2 in air at 37°C.

Cell growth assay

LNCaP cells (5 × 104 cells per well) were plated in 24-well plates (Costar, Corning Incorporated, Corning, NY); 24 h later, treatments were begun. Cells were treated with 0, 1, 5, 10 or 25 μM resveratrol (dimethylsulfoxide as vehicle) for 0–96 h, and the medium containing resveratrol was replaced every 24 h. Cell growth was analyzed using the sulforhodamine B assay as described previously.

For experiments using the synthetic androgen R1881 or 17β-estradiol, cells were switched to Media B [RPMI 1640 medium without phenol red (Invitrogen), 2 mM L-glutamine (Sigma), 100 U/ml penicillin and 100 μg/ml streptomycin with 10% charcoal–dextran-treated FBS (CDS; Hyclone, Logan, UT)] 24 h after plating to minimize the effect of serum steroid hormones. The cells were then incubated in Media B for an additional 24 h before the treatments were begun.

Article Produced By

Diet, Genomics and Immunology Laboratory, Beltsville Human Nutrition Research Center, Agriculture Research Service, United States Department of Agriculture, 10300 Baltimore Avenue, Building 307C, Room 132, Beltsville, MD. Laboratory of Cellular Regulation and Carcinogenesis, National Cancer Institutes, National Institutes of Health, Department of Nutrition and Food Science, University of Maryland, College Park, Department of Pharmaceutical Sciences and Pharmacology and Toxicology Graduate Program College of Pharmacy, Washington State University, Pullman, National Food Research Institute, Tsukuba, Ibaraki 305-8642, Japan. Nutritional Sciences Research Group, Division of Cancer Prevention, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. Biometrical Consulting Service, Beltsville Area, Agriculture Research Service, United States Department of Agriculture, Beltsville, MD 20705, USA. Division of Nutritional Sciences, University of Texas at Austin, Austin, TX 78712, USA. Department of Carcinogenesis, MD Anderson Cancer Center, Smithville, TX 78957, USA
 

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