PSA luciferase reporter gene assay

LNCaP cells were seeded into 24-well plates and were transiently transfected using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer’s instructions. LNCaP cells were cotransfected with 200 ng PSA-Luc reporter plasmid and 10 ng Renilla luciferase expression plasmid per well. Chemicals were added 24 h after transfection. The cells were harvested after another 24 h for a luciferase assay using the dual-luciferase reporter assay system (Promega, USA). The luciferase activities were normalized to the Renilla activity, which was cotransfected as an internal control. All of the assays were performed in triplicate, and the standard deviations were calculated accordingly.