Orotate phosphoribosyltransferase levels measured by a newly established enzyme-linked immunosorbent assay in gastric carcinoma.
A number of enzymes have been shown to be involved in the process of activation and/or degradation of 5-fluorouracil (5-FU), and are potential candidates for predicting chemosensitivity to 5-FU. Among these, orotate phosphoribosyltransferase (OPRT EC 2.4.2.10) is a key enzyme related to the first-step activation process of 5-FU and has been shown to be an important enzyme that helps to predict sensitivity to 5-FU and its related derivatives. We developed a new enzyme-linked immunosorbent assay (ELISA) to accurately assess intratumoral activity of OPRT. A new sandwich ELISA was established using anti-OPRT polyclonal antibodies obtained from the rabbit immunized with the recombinant human peptides of the OPRT molecule. OPRT levels were measured in eight human cancer xenografts and in 75 gastric cancer tissues using both a newly established ELISA and a conventional enzyme assay, using radiolabeled 5-FU as a substrate. There was a significant correlation between OPRT levels measured by this ELISA and OPRT enzyme activity the in eight human cancer xenografts (r2 = 0.782) and gastric carcinoma tissue (r2 = 0.617). The ELISA system for OPRT requires a minimal amount of carcinoma tissue, making it an easy-to-use assay system to predict sensitivity to 5-FU and its derivatives in gastric carcinoma. There was a significant correlation between tumor growth inhibition rates against the oral administration of oral-uracil/tegafur (UFT) and OPRT enzyme activity in the human cancer xenografts (r2 = 0.574). These results suggest that this newly developed sandwich ELISA system for the quantification of OPRT levels is technically simple, feasible and a useful tool to predict sensitivity to fluoropyrimidine-based anticancer chemotherapy in patients with gastric carcinoma and other cancers.
Sakurai
Yoichi
Y
Department of Surgery, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan. ysakurai@fujita-hu.ac.jp
Sakamoto
Kazuki
K
Sugimoto
Yoshikazu
Y
Yoshida
Ikuo
I
Masui
Toshihiko
T
Tonomura
Shuhei
S
Inaba
Kazuki
K
Shoji
Mitsutaka
M
Nakamura
Yasuko
Y
Uyama
Ichiro
I
Komori
Yoshiyuki
Y
Ochiai
Masahiro
M
Matsuura
Shiro
S
Tanaka
Hideyuki
H
Oka
Toshinori
T
Fukushima
Masakazu
M
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000378:metabolism; D019008:Drug Resistance, Neoplasm / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005472:Fluorouracil / Q000378:metabolism; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009962:Orotate Phosphoribosyltransferase / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000201:enzymology*; D014183:Transplantation, Heterologous
D000964:Antimetabolites, Antineoplastic; D009962:Orotate Phosphoribosyltransferase; D005472:Fluorouracil