13C NMR quantification of polyamine syntheses in rat prostate

Abstract

AbstractCurrently, many prostate cancer patients, detected through the prostate specific antigen test, harbor organ‐confined indolent disease that cannot be differentiated from aggressive cancer according to clinically and pathologically known measures. Spermine has been considered as an endogenous inhibitor for prostate‐confined cancer growth and its expression has shown correlation with prostate cancer growth rates. If established clinically, measurements of spermine bio‐synthesis rates in prostates may predict prostate cancer growth and patient outcomes. Using rat models, we tested the feasibility of quantifying spermine bio‐synthesis rates with 13C NMR. Male Copenhagen rats (10 weeks, n = 6) were injected with uniformly 13C‐labeled L‐ornithine HCl, and were sacrificed in pairs at 10, 30, and 60 min after injection. Another two rats were injected with saline and sacrificed at 30 min as controls. Prostates were harvested and extracted with perchloric acid and the neutralized solutions were examined by 13C NMR at 600 MHz. 13C NMR revealed measurable ornithine, as well as putrescine‐spermidine‐spermine syntheses in rat prostates, allowing polyamine bio‐synthetic and ornithine bio‐catabolic rates to be calculated. Our study demonstrated the feasibility of 13C NMR for measuring bio‐synthesis rates of ornithine to spermine enzymatic reactions in rat prostates. The current study established a foundation upon which future investigations of protocols that differentiate prostate cancer growth rates according to the measure of ornithine to spermine bio‐synthetic rates may be developed.