Abstract
The sensitivity to 5-fluorouracil (5-FU) has been reported to be associated with target molecule thymidylate synthase (TS), fluoropyrimidine-metabolising enzymes such as orotate phosphoribosyltransferase (OPRT), and dihydropyrimidine dehydrogenase (DPD). We performed an immunohistochemical study on the clinical significance of TS, OPRT, and DPD expression using 151 resected non-small-cell lung cancer (NSCLC) patients postoperatively treated with a combination of tegafur and uracil (UFT). Eighty-two carcinomas were TS-positive, 105 carcinomas were OPRT-positive, 68 carcinomas were DPD-positive. No correlation was observed in the HSCORE between the TS and OPRT expression (r=0.203), between the TS and DPD expression (r=0.098), or between the OPRT and DPD expression (r=0.074). Regarding the survival of NSCLC patients treated with UFT, the 5-year survival rate of patients with TS-negative tumours was significantly higher than that with TS-positive tumours (P=0.0133). The 5-year survival rate of patients with OPRT-positive stage II to III tumours was significantly higher than that with OPRT-negative stage II to III tumours (P=0.0145). In addition, the 5-year survival rate of patients with DPD-negative tumours was also significantly higher than that with DPD-positive tumours (P=0.0004). A Cox multivariate regression analysis revealed the TS status (hazard ratio 2.663; P=0.0003), OPRT status (hazard ratio 2.543; P=0.0005), and DPD status (hazard ratio 2.840; P<0.0001) to all be significant prognostic factors for the survival of resected NSCLC patients postoperatively treated with UFT.
Similar content being viewed by others
Main
Lung cancer is one of the most common cancers and the major cause of cancer-related death in North America, Europe, and Japan (Cersosimo, 2002). Non-small-cell lung cancer (NSCLC) comprises about 75% of all lung cancers. The majority of NSCLC patients have locally advanced stage III or metastatic stage IV disease at diagnosis. In addition, even in the early stage, some NSCLC patients may experience recurrence after surgery. Therefore, new therapeutic strategies are required to improve the clinical outcome of NSCLC patients.
Both experimental and clinical studies based on the molecular biology have revealed many molecules to affect various biological behaviors of malignant tumours including NSCLCs. To improve the treatment of NSCLC patients, it is important to design an optimal therapeutic strategy according to tumour biology (Huang et al, 2005). Therefore, the selection of effective adjuvant therapies based on biomarkers associated with tumour biology, such as made-to-order chemotherapy, is considered to be an effective strategy for NSCLC patients (Rosell et al, 2003; Vokes and Choy, 2003). Thus, the development of highly specific and responsive sensitivity tests for anticancer agents is important to be able to predict the response to an anticancer agent before using it for treatment. Recent experimental and clinical studies have reported several molecular targeted therapies, such as 5-fluorouracil (5-FU)-derived agents against thymidylate synthase (TS) (Rustum et al, 1997), Gefitinib or Erlotinib against epidermal growth factor receptor (Mitsudomi et al, 2005), and Bevacizumab against vascular endothelial growth factor-A (Johnson et al, 2004).
Among them, 5-FU-derived agents have been widely used in the treatment of various human cancers, including NSCLCs (Wada et al, 1996; Kato et al, 2004), colorectal cancers (Macdonald and Astrow, 2001), and gastric cancers (Takiuchi and Ajani, 1998), since its introduction into clinical practice more than 40 years ago. Clinical studies have reported their sensitivity to be associated with the target molecule TS (Rustum et al, 1997), fluoropyrimidine-metabolising enzymes such as orotate phosphoribosyltransferase (OPRT) (Peters et al, 1986), and the 5-FU-catabolitic enzyme such as dihydropyrimidine dehydrogenase (DPD) (Beck et al, 1994). However, there were only a few clinical studies on the simultaneous evaluations of these biomarkers to evaluate 5-FU sensitivity (Isshi et al, 2002; Fujii et al, 2003; Ichikawa et al, 2003a).
Therefore, we performed a clinical study on the clinical significance of TS, OPRT, and DPD expression for the treatment of resected NSCLC patients who were postoperatively treated with oral 5-FU derivatives, UFT (a combination of tegafur and uracil, Taiho Pharmaceutical Co., Tokyo, Japan) (Wada et al, 1996; Kato et al, 2004).
Materials and methods
Clinical characteristics of patients
From January 1995 to December 2000, consecutive NSCLC patients who underwent surgery and were postoperatively treated with UFT, at the Second Department of Surgery, Faculty of Medicine, Kagawa University, were studied. This study was approved by the institutional review board of Kagawa University (14-7, a clinical study of biological markers in NSCLCs) and signed informed consent was obtained from each patient. Tumour node metastasis staging designations were made according to the postsurgical pathological international staging system (Mountain, 1997). In total, 151 patients with lung cancer up to stage III were investigated (Table 1). They included 62 patients with stage I NSCLCs, 28 patients with stage II NSCLCs, and 61 patients with stage III NSCLCs. The patients' clinical records and histopathological diagnoses were fully documented. This report includes follow-up data as of 31 October 2005. The median follow-up period for all patients was 65.7 months.
Regarding the methods for a surgical resection, a pneumonectomy was performed in 14 patients with stage II to III NSCLCs. A lobectomy was performed in 123 patients; in 48 patients with stage I NSCLCs, in 26 patients with stage II NSCLCs, and in 49 patients with stage III NSCLCs. A segmentectomy was performed in four patients with stage I NSCLCs, and a wedge resection was performed in 10 patients with stage I NSCLCs. Platinum-based chemotherapy using mitomycin/vinblastin/cisplatin or carboplatin/paclitaxel was performed in 79 patients with stage II to III NSCLCs; neoadjuvant chemotherapy in 39 patients, and postoperative adjuvant chemotherapy in 40 patients with nodal metastases. Radiation therapy was performed in 24 patients; in 17 patients with T3 or T4 status and in seven patients with mediastinal lymph node metastases. The oral administration of UFT (300–400 mg day−1 body−1) was started within 1 month after surgery. UFT was administered for 2 years if there was no recurrence of carcinomas. In patients with recurrences, UFT was given until its oral administration became impossible.
Immunohistochemistry
Formalin-fixed paraffin-embedded tissue was cut into 4-μm sections and mounted on poly-L-lysine-coated slides. The sections were deparaffinised and rehydrated. The slides were then heated in a microwave for 10 min in a 10-μmol l−1 citrate buffer solution at pH 6.0, and cooled to room temperature for 20 min. After quenching the endogenous peroxidase activity with 0.3% H2O2 (in absolute methanol) for 30 min, the sections were treated for 2 h at room temperature with 5% bovine serum albumin to block nonspecific staining. Duplicate sections were incubated overnight with the primary specific antibodies detecting TS (rabbit IgG, diluted at 1 : 500), OPRT (rabbit IgG, diluted at 1 : 1000), and DPD (rabbit IgG, diluted at 1 : 500) (Huang et al, 2000). The slides were then incubated for 1 h with biotinylated anti-rabbit IgG (Vector Laboratories Inc., Burlingame, CA, USA). The sections were incubated with the avidin–biotin–peroxidase complex (Vector Laboratories Inc.) for 1 h, and antibody binding was visualised with 3,3′-diaminobenzidine tetrahydrochloride. Finally, the sections were lightly counterstained with Mayer's hematoxylin. The human colon cancer cell line DLD-1/FrUrd was used as a positive control for the staining of TS. Sections of resected lung tumours to express OPRT were used as positive controls for the staining of OPRT. The human pancreatic cancer cell line MIAPaCa-2 was used as a positive control for the staining of DPD. A rabbit IgG was used as a negative control during each run of immunohistochemical staining.
All of the immunostained sections were reviewed by two authors (J Nakano and M Ueno) who had no knowledge of the patients' clinical status. In cases of multiple areas of low intensity, five areas selected at random were scored. In sections where all of the staining appeared to be intense, one random field was selected. At least 200 tumour cells were scored per × 40 field. All sections were scored in a semiquantitative manner according to a previously described method, which reflects both the intensity and percentage of cells staining at each intensity (McCarty et al, 1986). Intensity was classified as 0 (no staining), +1 (weak staining), +2 (distinct staining), or +3 (very strong staining). A value designated as the ‘HSCORE’ was obtained for each slide by using the following algorithm: HSCORE=∑(I × PC), where I and PC represent the staining intensity and the percentage of cells that stain at each intensity, respectively. And the corresponding HSCOREs were calculated separately. Concerning TS expression, when the HSCORE of TS in a given specimen was ⩾30, the sample was classified as TS-positive, the same as reported previously (Huang et al, 2000). Concerning OPRT expression, the sample was classified as OPRT-positive when the HSCORE of OPRT in a given specimen was ⩾30, because the cutoff line of 30 showed the most significant association with a survival difference. Concerning DPD expression, when the HSCORE of DPD in a given specimen was ⩾50, the sample was classified as DPD-positive, the same as reported previously (Huang et al, 2000).
Statistical analysis
The statistical differences in the expression of each biological marker in relation to various clinical and pathological parameters were assessed by the χ2-test. Overall survival was defined as the time from treatment initiation (surgical resection, chemotherapy, or radiation) to the date of death from any cause. The Kaplan–Meier method was used to estimate the probability of overall survival as a function of time, and differences in the survival of subgroups of patients were compared by using Mantel's log-rank test. A multivariate analysis was performed using the Cox regression model to study the effects on survival (Cox, 1972). All P-values were based on two-tailed statistical analysis, and a P-value <0.05 was considered to indicate statistical significance.
Results
Clinical significance of intratumoural TS expression in NSCLCs treated with UFT
The intratumoural TS expression appeared in a cytoplasmic staining pattern (Figure 1A and 1B). Of the 151 NSCLCs we studied, 82 carcinomas (54.3%) were TS-positive, and 69 carcinomas (45.7%) were TS-negative (Table 2). The frequency of TS-positive tumours was significantly higher in squamous cell carcinomas than in adenocarcinomas (69.6 vs 42.9%, P=0.0063). However, there was no difference in the TS status according to tumour status, nodal status, pathological stage, tumour differentiation, or neoadjuvant chemotherapy.
Regarding the survival of NSCLC patients treated with UFT, the 5-year survival rate of patients with TS-negative tumours was significantly higher than that with TS-positive tumours (69.1 vs 44.7%, P=0.0133, Figure 2A). Regarding the pathological stage, the 5-year survival rate of patients with TS-negative stage I tumours was significantly higher than that with TS-positive stage I tumours (86.2 vs 63.2%, P=0.0426, Figure 2B). In addition, the 5-year survival rate of patients with TS-negative stage II to III tumours was also significantly higher than that with TS-positive stage II to III tumours (55.6 vs 27.9%, P=0.0259, Figure 2C).
Clinical significance of intratumoural OPRT expression in NSCLCs treated with UFT
The intratumoural OPRT expression appeared in a cytoplasmic staining pattern (Figure 1C and D). Of the 151 NSCLCs, 105 carcinomas (69.5%) were OPRT-positive, and 46 carcinomas (30.5%) were OPRT-negative (Table 2). There was no difference in the OPRT status according to tumour histology, tumour status, nodal status, pathological stage, tumour differentiation, or neoadjuvant chemotherapy.
Regarding the survival of NSCLC patients treated with UFT, there was no significant difference in the 5-year survival rates between patients with OPRT-positive tumours and patients with OPRT-negative tumours (60.6 vs 48.0%, Figure 3A). Regarding the pathological stage, there was also no significant difference in the 5-year survival rates of patients with stage I NSCLCs, according to the OPRT status (Figure 3B). However, among locally advanced stage II to III, the 5-year survival rate of patients with OPRT-positive tumours was significantly higher than that with OPRT-negative tumours (50.8 vs 25.0%, P=0.0145, Figure 3C).
Clinical significance of intratumoural DPD expression in NSCLCs treated with UFT
The intratumoural DPD expression appeared in a cytoplasmic staining pattern (Figure 1E and 1F). Of the 151 NSCLCs, 68 carcinomas (45.0%) were DPD-positive, and 83 carcinomas (55.0%) were DPD-negative (Table 2). There was no difference in the DPD status according to tumour histology, tumour status, nodal status, pathological stage, tumour differentiation, or neoadjuvant chemotherapy.
Regarding the survival of NSCLC patients treated with UFT, the 5-year survival rate of patients with DPD-negative tumours was significantly higher than that with DPD-positive tumours (66.6 vs 42.6%, P=0.0004, Figure 4A). Regarding the pathological stage, the 5-year survival rate of patients with DPD-negative stage I tumours was significantly higher than that with DPD-positive stage I tumours (79.8 vs 63.0%, P=0.0391, Figure 4B). In addition, the 5-year survival rate of patients with DPD-negative stage II to III tumours was also significantly higher than that with DPD-positive stage II to III tumours (52.4 vs 31.0%, P=0.0174, Figure 4C).
Prognostic factors of resected NSCLC patients treated with UFT
No correlation was observed in the HSCORE between the TS expression and OPRT expression (r=0.203), between the TS expression and DPD expression (r=0.098), or between the OPRT expression and DPD expression (r=0.074). A Cox multivariate regression analysis of the prognostic variables for resected NSCLC patients treated with UFT is shown in Table 3. Five variables, including the TS status (hazard ratio 2.663; P=0.0003), OPRT status (hazard ratio 2.534; P=0.0005), DPD status (hazard ratio 2.840; P<0.0001), pathological stage (hazard ratio 1.908; P<0.0001), and the methods of surgical resection (a pneumonectomy vs other methods)(hazard ratio 2.555; P=0.0242), were demonstrated to be significant prognostic factors for survival of resected NSCLC patients postoperatively treated with UFT.
Discussion
5-Fluorouracil is one of the most widely used antitumour chemothrapeutic agents for the treatment of a variety of human malignancies, especially in terms of the patient quality of life and economy (Wada et al, 1996). In fact, prospective randomised clinical studies have found the administration of UFT to be an effective adjuvant chemotherapy for NSCLC patients (Wada et al, 1996; Kato et al, 2004). On the other hand, many experimental and clinical studies have also revealed the 5-FU sensitivity to be affected by several biomarkers, such as its target molecule TS (Rustum et al, 1997), several fluoropyrimidine metabolising enzymes (Peters et al, 1986; Peters et al, 1991), and DPD, an enzyme of 5-FU catabolism (Beck et al, 1994).
First of all, regarding TS, its activity is necessary for cell proliferation because it catalyses the conversion of deoxyuridine-5′-monophosphate to deoxythymidine-5′-monophosphate, an essential step in DNA synthesis. After 5-FU is converted into 5-fluoro-2′-deoxyuridine 5′-monophosphate (FdUMP), this activated molecule forms a tight-binding complex with TS and 5,10-methylene-tetrahydrofolate, thus resulting in the inhibition of the TS activity (Rustum et al, 1997). Thymidylate synthase is therefore a target molecule of 5-FU. In addition, previous studies have reported that the regulation of the TS expression was affected by several mechanisms, including the polymorphism of the tandem repeated sequences in the promoter enhancer region of its gene and the genetic amplification of 18p (Horie et al, 1995; Hidaka et al, 2003).
In fact, many previous studies have reported the intratumoural TS expression to be associated with tumour proliferation (Navalgund et al, 1980; Nakagawa et al, 2004), the responsiveness to 5-FU (Johnston et al, 1995; Yeh et al, 1998). In addition, the overexpression of TS is reported to be associated with a poor prognosis of many cancer patients treated with 5-FU, including NSCLCs (Huang et al, 2000) and gastrointestinal cancers (Lenz et al, 1996). The present study in a relatively large number of patients treated with UFT also revealed that the survival of patients with TS-negative tumours was significantly better than that with TS-positive tumours. Furthermore, the significant differences in the survival of patients were observed in stage I cases as well as in locally advanced stage II to III cases. These results might be due to not only the responsiveness to 5-FU but also tumour proliferation (Nakagawa et al, 2004; Huang et al, 2005). In fact, recent studies have found that TS exhibits an oncogene-like activity (Rahman et al, 2004). In addition, the overexpression of TS itself is an independent factor for a poor prognosis in NSCLC patients (Huang et al, 2005).
Secondly, because 5-FU itself is a prodrug which requires its intracellular conversion, interindividual variation in the activity of fluoropyrimidine-metabolising enzymes can affect the activation of 5-FU. In general, phosphorylation is necessary to activate 5-FU by one or more of three pathways (Peters et al, 1986; Peters et al, 1991). Among them, OPRT is considered to be a major enzyme in the 5-FU conversion, directly to 5-fluorouridine 5′-monophosphate (FUMP) leading to the formation of FdUMP in the presence of 5-phosphoribosyl-1-pyrophosphate as a cofactor (Shirasaka et al, 1993). Experimental studies using cell lines have revealed a decreased expression and activity of OPRT to be associated with resistance to 5-FU (Chung et al, 2000; Murakami et al, 2000). Although the true mechanisms for the downregulation of OPRT expression in human cancers are still unclear, an experimental study has suggested the loss of 3q to be a genetic change responsible for the decreased OPRT activity and acquired resistance to 5-FU (Hidaka et al, 2003). Furthermore, the adenoviral-mediated transduction of OPRT gene was reported to result in a marked sensitisation of tumour cells to 5-FU cytotoxicity (Kanai et al, 1998).
Previous clinical studies also revealed the positive expression and activity of OPRT to be associated with the responsiveness to 5-FU in a variety of human cancers, including colorectal cancers (Isshi et al, 2002; Fujii et al, 2003; Ichikawa et al, 2003a; Matsuyama et al, 2006) and bladder carcinomas (Mizutani et al, 2004). However, these studies were performed using relatively small number of patients, and then were evaluated by biochemical assays for OPRT enzyme activity or reverse transcriptional PCR (RT–PCR) assays for OPRT gene expression. Biochemical assays are difficult to perform in clinical use, and they need a large volume of sample tissue. In the present study, we evaluated the intratumoural OPRT expression by immunohistochemistry using an anti-OPRT polyclonal antibody, obtained by immunising rabbits with the OPRT peptide. The immunochemical detection using this antibody was found to positively correlate with the OPRT enzyme activity (Sakamoto et al, 2005), and immunohistochemical assays are easy to perform in clinical use. Consequently, the present study demonstrated the positive expression of OPRT to be one of the significant factors for a good survival for resected NSCLC patients postoperatively treated with UFT, especially in advanced stage II to III. To our knowledge, the present study was the first clinical report using immunohistochemistry, to demonstrate the clinical significance of the OPRT expression in a relatively large number of cancer patients treated with UFT. In fact, a recent clinical study also revealed a positive OPRT activity to be associated with a better prognosis in resectable colorectal cancer patients treated with oral 5-FU (Ochiai et al, 2006).
On the other hand, regarding other pathways of 5-FU activation, thymidine phosphatase (TP) is considered to not be important because of the lack of its cofactor dRib-1-P at physiological concentrations (Barankiewicz and Henderson, 1977). Conversely, high TP concentration in conjunction with the normally low dRib-1-P drive the reaction in the opposite direction (Ackland and Peters, 1999). In fact, no difference was observed in the survivals of NSCLC patients treated with UFT according to the intratumoural TP expression in our pilot study by immunohistochemistry (data not shown).
Third, due to the fact that 80–90% of the administered 5-FU is degraded by DPD in humans, the first and rate-limiting enzyme of 5-FU catabolism, only a small part of the 5-FU is activated (Fischel et al, 1995). As a result, 5-FU degradation generally occurs in all tissues, including tumour tissues, but it reaches its highest level in the liver (Ho et al, 1986). In fact, a congenital deficiency of DPD can result in severe life-threatening toxicity when 5-FU is administered (Wei et al, 1996). In contrast, many experimental studies have demonstrated the intratumoural DPD activity to affect the resistance to 5-FU (Beck et al, 1994; Ishikawa et al, 1999). Previous clinical studies also found an overexpression of DPD in tumour cells to be associated with 5-FU resistance (Higashiyama et al, 2001) and a poor prognosis in many cancer patients treated with 5-FU, including NSCLCs (Huang et al, 2000; Nakagawa et al, 2002), colorectal cancers (Ichikawa et al, 2003b), and breast cancers (Horiguchi et al, 2004). Although several clinical studies reported DPD expression to not be associated with the survival of cancer patients treated with 5-FU (Ikeguchi et al, 2002; Kornmann et al, 2002; Hakamada et al, 2005), this discrepancy might be due to the different types of 5-FU administration such as intravenous infusion, the different antibody used, or the contamination of intratumoural stromal cells.
Based on these facts about the biomarkers associated with sensitivity to 5-FU, we thus considered a clinical study to simultaneously evaluate the expressions of TS, OPRT, and DPD, to be important for establishing the protocol of made-to-order chemotherapy for NSCLC patients. Therefore, we performed the present study using a relatively large number of patients treated with UFT. Consequently, the intratumoural expressions of TS, OPRT, and DPD were found to be independent of each other. Furthermore, a Cox multivariate analysis demonstrated all the intratumoural expressions of TS, OPRT, and DPD to be independent prognostic factors for resected NSCLC patients postoperatively treated with UFT. As a result, regarding the biomarkers associated with sensitivity to 5-FU, the survival of patients with TS-negative, OPRT-positive, and DPD-negative NSCLCs, which appeared in 12.6% (19 of 151) in the present study, was the highest among resected NSCLC patients treated with UFT.
Besides these biomarkers associated with sensitivity to 5-FU, the present study revealed that pathological stage and methods of surgical resection (a pneumonectomy vs other methods) were also independent prognostic factors for the resected NSCLC patients treated with UFT. Previous clinical studies also demonstrated that the survival of patients treated with a pneumonectomy is worse than that of patients treated with other surgical methods (Rocco et al, 1996; Ludwig et al 2005). On the other hand, regarding chemotherapy, most of patients with locally advanced stage II to III NSCLCs were treated with either neoadjuvant or postoperative adjuvant platinum-based chemotherapy using mitomycin/vinblastin/cisplatin or carboplatin/paclitaxel in the present study. However, we found no difference in the survivals of patients according to the platinum-based chemotherapy (data not shown). In order to clarify the clinical significance of the platinum-based chemotherapy for NSCLC patients, a randomised prospective study should thus be performed. Furthermore, among the locally advanced stage II to III cases, no difference was observed in the survivals of patients according to radiotherapy in the present study (data not shown). In fact, previous clinical studies also reported that radiotherapy itself has less survival benefit for NSCLC patients (Dautzenberg et al, 1999).
The efficacy of UFT is still lower in NSCLCs than in gastrointestinal cancers. For one reason, NSCLCs are human tumours with a high expression of DPD because DPD-positive tumours occurred in 45% of NSCLCs in the present study. Therefore, it is important to inhibit the DPD activity to improve the responsiveness to 5-FU against DPD-positive NSCLCs. Recently, S-1 (a combination of tegafur, gimeracil, and oteracil potassium, Taiho) has been developed for clinical use (Chu et al, 2004; Ichinose et al, 2004). Gimeracil used in S-1 is a stronger inhibitor for the intratumoural DPD activity than uracil used in UFT. As a result, S-1 might be more effective for patients with TS-negative, OPRT-positive, and DPD-positive NSCLCs, which occurred in 15.9% (24 of 151) in the present study. Therefore, patients with TS-negative and OPRT-positive NSCLCs, which appeared in 28.5% (43 of 151) in the present study, could be successfully treated by S-1 without evaluating the intratumoural DPD activity. In order to develop and simplify the protocol of made-to-order chemotherapy, a further randomised prospective study using S-1 should thus be performed to evaluate this possibility.
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Ackland SP, Peters GJ (1999) Thymidine phosphorylase: its role in sensitivity and resistance to anticancer drugs. Drug Resist Updat 2: 205–214
Barankiewicz J, Henderson JF (1977) Ribose 1-phosphate metabolism in Ehrlich ascites tumor cells in vitro. Biochim Biophys Acta 479: 371–377
Beck A, Etienne MC, Cheradame S, Fischel JL, Formento P, Renee N, Milano G (1994) A role for dihydropyrimidine dehydrogenase and thymidylate synthase in tumour sensitivity to fluorouracil. Eur J Cancer 30A: 1517–1522
Cersosimo RJ (2002) Lung cancer: a review. Am J Health Syst Pharm 59: 611–642
Chu QS, Hammond LA, Schwartz G, Ochoa L, Rha SY, Denis L, Molpus K, Roedig B, Letrent SP, Damle B, DeCillis AP, Rowinsky EK (2004) Phase I and pharmacokinetic study of the oral fluoropyrimidine S-1 on a once-daily-for-28-day schedule in patients with advanced malignancies. Clin Cancer Res 10: 4913–4921
Chung YM, Park S, Park JK, Kim Y, Kang Y, Yoo YD (2000) Establishment and characterization of 5-fluorouracil-resistant gastric cancer cells. Cancer Lett 159: 95–101
Cox DR (1972) Regression models and life-tables. J R Stat Soc B 34: 187–202
Dautzenberg B, Arriagada R, Chammard AB, Jarema A, Mezzetti M, Mattson K, Lagrange J, Pechoux C, Lebeau B, Chastang C (1999) A controlled study of postoperative radiotherapy for patients with completely resected nonsmall cell lung carcinoma. Cancer 86: 265–273
Fischel JL, Etienne MC, Spector T, Formento P, Renee N, Milano G (1995) Dihydropyrimidine dehydrogenase: a tumoral target for fluorouracil modulation. Clin Cancer Res 1: 991–996
Fujii R, Seshimo A, 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
Hakamada Y, Tsuchida A, Arima M, Kubouchi T, Tokita H, Ota D, Kaise H, Aoki T, Kusama M, Aoki T (2005) Prognostic predictors in breast cancer patients with postoperative 5-fluorouracil-based chemotherapy. Int J Mol Med 16: 309–314
Hidaka S, Yasutake T, Fukushima M, Yano H, Haseba M, Tsuji T, Sawai T, Yamaguchi H, Nakagoe T, Ayabe H, Tagawa Y (2003) Chromosomal imbalances associated with acquired resistance to fluoropyrimidines in human colorectal cancer cells. Eur J Cancer 39: 975–980
Higashiyama M, Kodama K, Yokouchi H, Takami K, Fukushima M, Minamigawa K, Takano T, Kobayashi H (2001) Thymidylate synthase and dihydropyrimidine dehydrogenase activities in non-small cell lung cancer tissues: relationship with in vitro sensitivity to 5-fluorouracil. Lung Cancer 34: 407–416
Ho DH, Townsend L, Luna MA, Bodey GP (1986) Distribution and inhibition of dihydrouracil dehydrogenase activities in human tissues using 5-fluorouracil as a substrate. Anticancer Res 6: 781–784
Horie N, Aiba H, Oguro K, Hojo H, Takeishi K (1995) Functional analysis and DNA polymorphism of the tandemly repeated sequences in the 5′-terminal regulatory region of the human gene for thymidylate synthase. Cell Struct Funct 20: 191–197
Horiguchi J, Yoshida T, Koibuchi Y, Iijima K, Ninomiya J, Takei H, Yokoe T, Iino Y, Morishita Y (2004) DPD activity and immunohistochemical DPD expression in human breast cancer. Oncol Rep 11: 65–72
Huang CL, Yokomise H, Kobayashi S, Fukushima M, Hitomi S, Wada H (2000) Intratumoral expression of thymidylate synthase and dihydropyrimidine dehydrogenase in non-small cell lung cancer patients treated with 5-FU-based chemotherapy. Int J Oncol 17: 47–54
Huang C, Liu D, Masuya D, Nakashima T, Kameyama K, Ishikawa S, Ueno M, Haba R, Yokomise H (2005) Clinical application of biological markers for treatments of respectable non-small-cell lung cancers. Br J Cancer 92: 1231–1239
Ichikawa W, Uetake H, Shirota Y, Yamada H, Takahashi T, Nihei Z, Sugihara K, Sasaki Y, Hirayama R (2003a) Both gene expression for orotate phosphoribosyltransferase and its ratio to dihydropyrimidine dehydrogenase influence outcome following fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. Br J Cancer 89: 1486–1492
Ichikawa W, Uetake H, Shirota Y, Yamada H, Nishi N, Nihei Z, Sugihara K, Hirayama R (2003b) Combination of dihydropyrimidine dehydrogenase and thymidylate synthase gene expressions in primary tumors as predictive parameters for the efficacy of fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. Clin Cancer Res 9: 786–791
Ichinose Y, Yoshimori K, Sakai H, Nakai Y, Sugiura T, Kawahara M, Niitani H (2004) S-1 plus cisplatin combination chemotherapy in patients with advanced non-small cell lung cancer: a multi-institutional phase II trial. Clin Cancer Res 10: 7860–7864
Ikeguchi M, Makino M, Kaibara N (2002) Thymidine phosphorylase and dihydropyrimidine dehydrogenase activity in colorectal carcinoma and patients prognosis. Langenbecks Arch Surg 387: 240–245
Isshi K, Sakuyama T, Gen T, Nakamura Y, Kuroda T, Katuyama T, 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
Ishikawa Y, Kubota T, Otani Y, Watanabe M, Teramoto T, Kumai K, Kitajima M, Takechi T, Okabe H, Fukushima M (1999) Dihydropyrimidine dehydrogenase activity and messenger RNA level may be related to the antitumor effect of 5-fluorouracil on human tumor xenografts in nude mice. Clin Cancer Res 5: 883–889
Johnson DH, Fehrenbacher L, Novotny WF, Herbst RS, Nemunaitis JJ, Jablons DM, Langer CJ, DeVore III RF, Gaudreault J, Damico LA, Holmgren E, Kabbinavar F (2004) Randomized phase II trial comparing bevacizumab plus carboplatin and paclitaxel with carboplatin and paclitaxel alone in previously untreated locally advanced or metastatic non-small-cell lung cancer. J Clin Oncol 22: 2184–2191
Johnston PG, Lenz HJ, Leichman CG, Danenberg KD, Allegra CJ, Danenberg PV, Leichman L (1995) Thymidylate synthase gene and protein expression correlate and are associated with response to 5-fluorouracil in human colorectal and gastric tumors. Cancer Res 55: 1407–1412
Kanai F, Kawakami T, Hamada H, Sadata A, Yoshida Y, Tanaka T, Ohashi M, Tateishi K, Shiratori Y, Omata M (1998) Adenovirus-mediated transduction of Escherichia coli uracil phosphoribosyltransferase gene sensitizes cancer cells to low concentrations of 5-fluorouracil. Cancer Res 58: 1946–1951
Kato H, Ichinose Y, Ohta M, Hata E, Tsubota N, Tada H, Watanabe Y, Wada H, Tsuboi M, Hamajima N, Ohta M, Japan Lung Cancer Research Group on Postsurgical Adjuvant Chemotherapy (2004) A randomized trial of adjuvant chemotherapy with uracil-tegafur for adenocarcinoma of the lung. N Engl J Med 350: 1713–1721
Kornmann M, Link KH, Galuba I, Ott K, Schwabe W, Hausler P, Scholz P, Strater J, Polat S, Leibl B, Kettner E, Schlichting C, Baumann W, Schramm H, Hecker U, Ridwelski K, Vogt JH, Zerbian KU, Schutze F, Kreuser ED, Behnke D, Beger HG (2002) Association of time to recurrence with thymidylate synthase and dihydropyrimidine dehydrogenase mRNA expression in stage II and III colorectal cancer. J Gastrointest Surg 6: 331–337
Lenz HJ, Leichman CG, Danenberg KD, Danenberg PV, Groshen S, Cohen H, Laine L, Crookes P, Silberman H, Baranda J, Garcia Y, Li J, Leichman L (1996) Thymidylate synthase mRNA level in adenocarcinoma of the stomach: a predictor for primary tumor response and overall survival. J Clin Oncol 14: 176–182
Ludwig C, Stoelben E, Olschewski M, Hasse J (2005) Comparison of morbidity, 30-day mortality, and long-term survival after pneumonectomy and sleeve lobectomy for non-small cell lung carcinoma. Ann Thorac Surg 79: 968–973
Macdonald JS, Astrow AB (2001) Adjuvant therapy of colon cancer. Semin Oncol 28: 30–40
Matsuyama R, Togo S, Shimizu D, Momiyama N, Ishikawa T, Ichikawa Y, Endo I, Kunisaki C, Suzuki H, Hayasizaki Y, Shimada H (2006) Predicting 5-fluorouracil chemosensitivity of liver metastases from colorectal cancer using primary tumor specimens: Three-gene expression model predicts clinical response. Int J Cancer 119: 406–413
McCarty Jr KS, Szabo E, Flowers JL, Cox EB, Leight GS, Miller L, Konrath J, Soper JT, Budwit DA, Creasman WT, Seigler HF, McCarty Sr KS (1986) Use of a monoclonal anti-estrogen receptor antibody in the immunohistochemical evaluation of human tumors. Cancer Res 46: 4244s–4248s
Mitsudomi T, Kosaka T, Endoh H, Horio Y, Hida T, Mori S, Hatooka S, Shinoda M, Takahashi T, Yatabe Y (2005) Mutations of the epidermal growth factor receptor gene predict prolonged survival after gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence. J Clin Oncol 23: 2513–2520
Mizutani Y, Wada H, Fukushima M, Yoshida O, Nakanishi H, Li YN, Miki T (2004) Prognostic significance of orotate phosphoribosyltransferase activity in bladder carcinoma. Cancer 100: 723–731
Mountain CF (1997) Revisions in the international system for staging lung cancer. Chest 111: 1710–1717
Murakami Y, Kazuno H, Emura T, Tsujimoto H, Suzuki N, Fukushima M (2000) Different mechanisms of acquired resistance to fluorinated pyrimidines in human colorectal cancer cells. Int J Oncol 17: 277–283
Nakagawa T, Tanaka F, Takata T, Matsuoka K, Miyahara R, Otake Y, Yanagihara K, Fukushima M, Wada H (2002) Predictive value of dihydropyrimidine dehydrogenase expression in tumor tissue, regarding the efficacy of postoperatively administered UFT (Tegafur+Uracil) in patients with p-stage I nonsmall-cell lung cancer. J Surg Oncol 81: 87–92
Nakagawa T, Otake Y, Yanagihara K, Miyahara R, Ishikawa S, Fukushima M, Wada H, Tanaka F (2004) Expression of thymidylate synthase is correlated with proliferative activity in non-small cell lung cancer (NSCLC). Lung Cancer 43: 145–149
Navalgund LG, Rossana C, Muench AJ, Johnson LF (1980) Cell cycle regulation of thymidylate synthetase gene expression in cultured mouse fibroblasts. J Biol Chem 255: 7386–7390
Ochiai T, Nishimura K, Noguchi H, Kitajima M, Tsukada A, Watanabe E, Nagaoka I, Futagawa S (2006) Prognostic impact of orotate phosphoribosyl transferase among 5-fuorouracil metabolic enzymes in resectable colorectal cancers treated by oral 5-fluorouracil-based adjuvant chemotherapy. Int J Cancer 118: 3084–3088
Peters GJ, Laurensse E, Leyva A, Lankelma J, Pinedo HM (1986) Sensitivity of human, murine, and rat cells to 5-fluorouracil and 5′-deoxy-5-fluorouridine in relation to drug-metabolizing enzymes. Cancer Res 46: 20–28
Peters GJ, van Groeningen CJ, Laurensse EJ, Pinedo HM (1991) A comparison of 5-fluorouracil metabolism in human colorectal cancer and colon mucosa. Cancer 68: 1903–1909
Rahman L, Voeller D, Rahman M, Lipkowitz S, Allegra C, Barrett JC, Kaye FJ, Zajac-Kaye M (2004) Thymidylate synthase as an oncogene: a novel role for an essential DNA synthesis enzyme. Cancer Cell 5: 341–351
Rocco PM, Antkowiak JG, Takita H, Urschel JD (1996) Long-term outcome after pneumonectomy for nonsmall cell lung cancer. J Surg Oncol 61: 278–280
Rosell R, Taron M, Alberola V, Massuti B, Felip E (2003) Genetic testing for chemotherapy in non-small cell lung cancer. Lung Cancer 41: S97–S102
Rustum YM, Harstrick A, Cao S, Vanhoefer U, Yin MB, Wilke H, Seeber S (1997) Thymidylate synthase inhibitors in cancer therapy: direct and indirect inhibitors. J Clin Oncol 15: 389–400
Sakamoto K, Sugimoto Y, Miyadera K, Oka T, Fukushima M (2005) Preparation of anti-orotate phosphoribosyltransferase antibody and its application to immunochemical detection in human tumor cells. Int J Mol Med 16: 245–249
Shirasaka T, Shimamoto Y, Fukushima M (1993) Inhibition by oxonic acid of gastrointestinal toxicity of 5-fluorouracil without loss of its antitumor activity in rats. Cancer Res 53: 4004–4009
Takiuchi H, Ajani JA (1998) Uracil-tegafur in gastric carcinoma: a comprehensive review. J Clin Oncol 16: 2877–2885
Vokes EE, Choy H (2003) Targeted therapies for stage III non-small cell lung cancer: integration in the combined modality setting. Lung Cancer 41: S115–S121
Wada H, Hitomi S, Teramatsu T (1996) Adjuvant chemotherapy after complete resection in non-small-cell lung cancer. West Japan Study Group for Lung Cancer Surgery. J Clin Oncol 14: 1048–1054
Wei X, McLeod HL, McMurrough J, Gonzalez FJ, Fernandez-Salguero P (1996) Molecular basis of the human dihydropyrimidine dehydrogenase deficiency and 5-fluorouracil toxicity. J Clin Invest 98: 610–615
Yeh KH, Shun CT, Chen CL, Lin JT, Lee WJ, Lee PH, Chen YC, Cheng AL (1998) High expression of thymidylate synthase is associated with the drug resistance of gastric carcinoma to high dose 5-fluorouracil-based systemic chemotherapy. Cancer 82: 1626–1631
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Nakano, J., Huang, C., Liu, D. et al. Evaluations of biomarkers associated with 5-FU sensitivity for non-small-cell lung cancer patients postoperatively treated with UFT. Br J Cancer 95, 607–615 (2006). https://doi.org/10.1038/sj.bjc.6603297
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6603297
Keywords
This article is cited by
-
5-Fluorouracil Loaded Chitosan–PVA/Na+MMT Nanocomposite Films for Drug Release and Antimicrobial Activity
Nano-Micro Letters (2016)
-
Cyclooxygenase-2 expression is a prognostic biomarker for non-small cell lung cancer patients treated with adjuvant platinum-based chemotherapy
World Journal of Surgical Oncology (2015)
-
MicroRNA-433 negatively regulates the expression of thymidylate synthase (TYMS) responsible for 5-fluorouracil sensitivity in HeLa cells
BMC Cancer (2013)
-
A phase 1 study of TRC102, an inhibitor of base excision repair, and pemetrexed in patients with advanced solid tumors
Investigational New Drugs (2013)
-
Thymidylate synthase expression is closely associated with outcome in patients with pulmonary adenocarcinoma
Medical Oncology (2012)