Quantitative Analysis of the Enzymes Associated with 5-Fluorouracil Metabolism in Prostate Cancer Biopsies

Abstract

Orotate phosphoribosyl transferase (OPRT) is the initial enzyme of 5-FU activation, in which 5-FU is converted to 5-fluorouridinemonophosphate. Dihydropyrimidine dehydrogenase (DPD) is a degrading enzyme that catabolizes 5-FU. In this study, we investigated the expression of these enzymes in normal prostate gland (NP), hormone-sensitive prostate cancer (HSPC), and hormone-refractory prostate cancer (HRPC). The prostatic tissue specimens were obtained from patients who had undergone prostate needle biopsies without any treatments or with PSA failure after initial androgen deprivation. The tissue samples derived from formalin-fixed, paraffin-embedded (FFPE) sections were prepared by laser-capture microdissection, and from them RNA was extracted. The levels of OPRT and DPD mRNA expression were examined by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The level of OPRT mRNA expression in the HSPC or the HRPC specimens was significantly higher than that in the NP specimens. There was a significant correlation between OPRT mRNA expression levels and the tumor pathological grade. Furthermore, the OPRT/ DPD expression ratio, a powerful predictive factor to evaluate 5-FU sensitivity, in the HRPC group was significantly higher than that in the low-grade HSPC group. Thus, the quantitative evaluation for these enzymes based on phosphorylation of 5-FU may be an effective option for some prostate cancer patients, particularly HRPC group.

The Strange Case of Orotic Acid: The Different Expression of Pyrimidines Biosynthesis in Healthy Males and Females

“…The complete pathway of OA biosynthesis is reported in Figure 1. The step (5), represented in Figure 1, is directly involved in the metabolism of 5-FU because this anticancer drug is a competitive substrate of OPRTase [4]. In particular, the transferase activity of OPRTase multicomplex enzyme is inhibited by 5-FU at 59% level of control [5].…”