Sex Hormone Binding Globulin Modifies Testosterone Action and Metabolism in Prostate Cancer Cells

Figure 2

Effect of SHBG on testosterone uptake and metabolism by LNCaP cells measured by Liquid Chromatography/Tandem Mass (LC MSMS) Spectrometry. LNCaP cells were treated with 25 nM Te ± 125 nM SHBG (S) for 1, 4, and 24 hours. (a) Cellular Te and (b) total Te in culture medium and cell lysates (undigested). (c) Measurement of unconjugated (black) and glucuronidated (grey) Te in cell culture medium. Each medium sample was divided into two equal volumes and digested with or without 200 units/mL β-glucuronidase in potassium phosphate buffer (pH 6.9) for 24 hours at 37°C to revert glucuronidated testosterone to unconjugated testosterone. All measurements are expressed as a percentage of Te in the original uptake medium.

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