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

  • Tomoaki Tanaka
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 755)

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.

Key words

Prostate cancer 5-FU Orotate phosphoribosyl transferase Dihydropyrimidine dehydrogenase Laser-cutting microdissection RT-PCR 

References

  1. 1.
    Longley, D. B., Harkin, D. P., and Johnston, P. G. (2003) 5-fluorouracil: mechanisms of action and clinical strategies, Nat Rev Cancer 3, 330–338.CrossRefGoogle Scholar
  2. 2.
    Miyake, H., Hara, I., Yamazaki, H., and Eto, H. (2005) Clinical outcome of oral uracil/tegafur (UFT) therapy for patients with hormone refractory prostate cancer, Oncol Rep 14, 673–676.Google Scholar
  3. 3.
    Bhandari, M. S., Pienta, K. J., Fardig, J., Olson, K., and Smith, D. C. (2006) Phase II trial of oral uracil/tegafur plus leucovorin in patients with hormone-refractory prostate carcinoma, Cancer 106, 1715–1721.CrossRefGoogle Scholar
  4. 4.
    Hoshi, S., Ohyama, C., Hagisawa, S., Ono, K., Satoh, M., Saito, S., Fukuzaki, A., and Arai, Y. (2003) Complete regression of bone metastases on super bone scan, by low-dose cisplatin, UFT, diethylstilbestrol diphosphate, and dexamethasone in a patient with hormone-refractory prostate cancer, Int J Clin Oncol 8, 118–120.CrossRefGoogle Scholar
  5. 5.
    Kubota, T. (2003) 5-fluorouracil and dihydropyrimidine dehydrogenase, Int J Clin Oncol 8, 127–131.CrossRefGoogle Scholar
  6. 6.
    Isshi, K., Sakuyama, T., Gen, T., Nakamura, Y., Kuroda, T., Katuyama, T., and Maekawa, Y. (2002) Predicting 5-FU sensitivity using human colorectal cancer specimens: comparison of tumor dihydropyrimidine dehydrogenase and orotate phosphoribosyl transferase activities with in vitro chemosensitivity to 5-FU, Int J Clin Oncol 7, 335–342.CrossRefGoogle Scholar
  7. 7.
    Oguri, T., Achiwa, H., Bessho, Y., Muramatsu, H., Maeda, H., Niimi, T., Sato, S., and Ueda, R. (2005) The role of thymidylate synthase and dihydropyrimidine dehydrogenase in resistance to 5-fluorouracil in human lung cancer cells, Lung Cancer 49, 345–351.CrossRefGoogle Scholar
  8. 8.
    Sakamoto, E., Nagase, H., Kobunai, T., Oie, S., Oka, T., Fukushima, M., and Oka, T. (2007) Orotate phosphoribosyltransferase expression level in tumors is a potential determinant of the efficacy of 5-fluorouracil, Biochem Biophys Res Commun 363, 216–222.CrossRefGoogle Scholar
  9. 9.
    Wada, Y., Yoshida, K., Suzuki, T., Mizuiri, H., Konishi, K., Ukon, K., Tanabe, K., Sakata, Y., and Fukushima, M. (2006) Synergistic effects of docetaxel and S-1 by modulating the expression of metabolic enzymes of 5-fluorouracil in human gastric cancer cell lines, Int J Cancer 119, 783–791.CrossRefGoogle Scholar
  10. 10.
    Fujii, R., Seshimo, A., and Kameoka, S. (2003) Relationships between the expression of thymidylate synthase, dihydropyrimidine dehydrogenase, and orotate phosphoribosyltransferase and cell proliferative activity and 5-fluorouracil sensitivity in colorectal carcinoma, Int J Clin Oncol 8, 72–78.CrossRefGoogle Scholar
  11. 11.
    Katsumata, K., Tomioka, H., Sumi, T., Yamashita, S., Takagi, M., Kato, F., Nakamura, R., Koyanagi, Y., Aoki, T., and Kato, K. (2003) Correlation between clinicopathologic factors and kinetics of metabolic enzymes for 5-fluorouracil given to patients with colon carcinoma by two different dosage regimens, Cancer Chemother Pharmacol 51, 155–160.Google Scholar
  12. 12.
    Birtle, A. J., Newby, J. C., and Harland, S. J. (2004) Epirubicin carboplatin and 5-fluorouracil (ECarboF) chemotherapy in metastatic hormone refractory prostate cancer, Br J Cancer 91, 1472–1476.Google Scholar
  13. 13.
    Droz, J. P., Muracciole, X., Mottet, N., Ould Kaci, M., Vannetzel, J. M., Albin, N., Culine, S., Rodier, J. M., Misset, J. L., Mackenzie, S., Cvitkovic, E., and Benoit, G. (2003) Phase II study of oxaliplatin versus oxaliplatin combined with infusional 5-fluorouracil in hormone refractory metastatic prostate cancer patients, Ann Oncol 14, 1291–1298.CrossRefGoogle Scholar
  14. 14.
    Tanaka, T., Kawashima, H., Matsumura, K., Yamashita-Hosono, T., Yoshimura, R., Kuratsukuri, K., Harimoto, K., and Nakatani, T. (2009) Overexpression of orotate phosphoribosyl transferase in hormone-refractory prostate cancer, Oncol Rep 21, 33–37.CrossRefGoogle Scholar
  15. 15.
    Lord, R. V., Salonga, D., Danenberg, K. D., Peters, J. H., DeMeester, T. R., Park, J. M., Johansson, J., Skinner, K. A., Chandrasoma, P., DeMeester, S. R., Bremner, C. G., Tsai, P. I., and Danenberg, P. V. (2000) Telomerase reverse transcriptase expression is increased early in the Barrett’s metaplasia, dysplasia, adenocarcinoma sequence, J Gastrointest Surg 4, 135–142.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of UrologyOsaka City University Graduate School of MedicineOsakaJapan

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