Cell Culture and Animal Models

The PC-3 human prostate carcinoma cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD). The cells were grown in Roswell Park Memorial Institute (RPMI) 1640 Medium supplemented with 10% FBS, penicillin (100 IU/mL), and streptomycin (100 mg/mL) (Invitrogen, Carlsbad, CA) and cultured at 37 °C in a humidified atmosphere containing 5% CO₂. Cells were passaged three times per week.

All animal studies were conducted according to the principles and procedures outlined in the Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of the Clinical Center, National Institutes of Health (NIH). Female BALB/c mice (age, 5–7 weeks; weights of 18–20 g) and female athymic nude/nude mice (age, 5–7 weeks; weights of 18–20 g) were purchased from Harlan Laboratories.

The PC-3 tumor models were generated by subcutaneous injection of 5 × 10⁶ cells into the right shoulder of nude mice. Tumor sizes were measured using a digital caliper. Tumor volume (mm³) was calculated according to the formula 0.5 × length × width². The mice were subjected to micro PET studies when the tumor volume reached 100–300 mm³ (3–4 weeks after inoculation).