Previous studies have reported that the expressions of specific proteins may predict the efficacy of chemotherapy agents for non-small cell lung cancer (NSCLC) patients. The present study evaluated the expression of proteins hypothesized to be associated with the effect of chemotherapeutic agents in 38 NSCLC patients with pathological stage II and IIIA. The subjects received carboplatin plus paclitaxel (CP) or S-1 as adjuvant chemotherapy following complete resection. The protein expressions evaluated were those of thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phsphoribosyltransferase (OPRT), which were suspected to be associated with the effect of S-1 agents, excision repair cross-complementation group 1 (ERCC1), which was suspected to be associated with the effect of platinum-based agents, and class III β-tubulin (TUBB3), which was suspected to be associated with the effect of taxane-based agents. The positive rate of TS was 55.3% (n=21/38), DPD was 57.9% (n=22/38), OPRT was 42.1% (n=16/38), ERCC1 was 47.4% (n=18/38) and TUBB3 was 44.7% (n=17/38). Among the patients who received S-1 adjuvant chemotherapy, TS-negative cases demonstrated a significantly better disease-free survival than positive cases. Thus, TS protein expression may have been a factor that predicted the effect of S-1 agent as adjuvant chemotherapy.
non-small cell lung canceradjuvant chemotherapyprotein expressionthymidylate synthasedihydropyrimidine dehydrogenaseorotate phsphoribosyltransferaseexcision repair cross-complementation group 1class III β-tubulinIntroduction
The mortality rates of patients with advanced non-small cell lung cancer (NSCLC) remain high (1). To improve this poor prognosis, several adjuvant chemotherapies have been administered in patients with completely resected NSCLC, but the improvement of the survival rate is not ideal, and patients sometimes struggle with adverse effects, like nausea, neutropenia, and fatigue (2–10). Ideally, we would be able to predict the effects of chemotherapeutic agents and regimens for patients who received chemotherapy, especially for postoperative adjuvant chemotherapy, because whether or not adjuvant chemotherapy reduces the rate of recurrence is unclear. Even with cytotoxic anticancer drugs, the predictive factors of the therapeutic effect would ideally be revealed in a manner similar to that observed for molecular targeted therapy (11–13).
Recently, the expression of some proteins has been reported as a predictor of the efficacy of cytotoxic chemotherapeutic agents. Excision repair cross-complementation group 1 (ERCC1) is a DNA repair gene in the nucleotide excision repair pathway that is activated when platinum-based chemotherapeutic agents form DNA adducts (14). High ERCC1 expression in several cancers has been reported in association with resistance to platinum-based treatment (15–17). Class III β-tubulin (TUBB3) is a major component of the microtubules that are targeted by taxane-based agents, which exert their growth inhibitory effects through the inhibition of microtubule dynamics, resulting in the growth arrest of tumor cells at the G2-M phase (18). High expression of TUBB3 has been reported in association with resistance to taxane-based treatment in human cancers (19–21). Thymidylate synthase (TS) is an enzyme that generates deoxythymidine monophosphate, which is subsequently phosphorylated to thymidine triphosphate for use in DNA synthesis and repair. High expression of TS has been reported in association with fluorouracil (5FU)-based chemotherapy (including S-1 agent) resistance in various cancers (22–24). Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in degrading 5-FU to 2-fluoro-β-alanine (25), and high expression of DPD has been reported in association with resistance to 5-FU-based chemotherapies (26–28). Orotate phosphoribosyltransferase (OPRT) is an enzyme involved in pyrimidine biosynthesis and contributes to the conversion of 5-FU into fdUMP, an active form of 5-FU. Low expression of OPRT has been reported in association with resistance to 5-FU-based chemotherapies (29,30).
In this study, we investigated the expression of several proteins in completely resected NSCLC patients who received carboplatin plus paclitaxel (CP) or S-1 regimen as adjuvant chemotherapy.
Patients and methodsPatients
A multicenter randomized feasibility study of CP vs. S-1 in patients with locally advanced completely resected NSCLC was conducted. Forty patients underwent complete resection and were diagnosed with pathological stage II or IIIA NSCLC (the 7th edition of the Tumor-Node-Metastasis classification) (31) at Nagoya City University Hospital (Nagoya, Japan) and its affiliated hospitals between January 2008 and December 2013.
Written informed consent was obtained from all patients, and the study protocol was approved by the Institutional Review Board of each participating institution (Nagoya City University Hospital No. 45-13-0020). This study was registered on the UMIN Clinical Trial database (ID:000001510). We have reported on details of this study (32). In this paper, we evaluated the relationships between the protein expression and the prognosis of patients who received adjuvant chemotherapy after complete surgical resection. The randomization was performed centrally at the Department of Oncology, Immunology and Surgery, Nagoya City University Graduate School of Medical Sciences (Nagoya, Japan).
Design of the study and treatment schedule
The patients were randomly assigned either to arm A (21 cases) receiving CP bi-weekly or to arm B (19 cases) receiving S-1. Among the 40 patients, two patients assigned to arm A could not continue the adjuvant chemotherapy because of a Grade 4 allergic reaction (anaphylactic shock) during the first cycle of paclitaxel infusion. We excluded these two patients from this additional study and investigated the 38 patients who received adjuvant chemotherapy over two courses.
The infusing dosage of paclitaxel was 120 mg m−2 on days 1 and 15. Carboplatin at an area under the curve (AUC 3) dose was also administered on days 1 and 15. The patients received adjuvant chemotherapy with carboplatin plus paclitaxel every four weeks for up to four cycles. Calvert's formula was used to calculate the dose of the AUC for carboplatin (33), while the creatinine clearance was determined with the Jellifie formula (34). The dosage of S-1 was established as follows: patients with a body surface area (BSA) <1.25 m2 received 40 mg twice a day (80 mg/day), those with BSA of ≥1.25 m2 but <1.5 m2 received 50 mg twice a day (100 mg/day), and those with a BSA ≥1.5 m2 received 60 mg twice a day (120 mg/day). S-1 was administered for two weeks followed by a one-week rest period for up to one year. Both arms A and B continued on the above prescription unless any evidence for relapse, other malignancies, or severe adverse events were identified.
Recurrence was diagnosed on the basis of imaging study findings. Chest and abdominal computed tomography and positron emission tomography plus head magnetic resonance imaging were performed at 6- and 12-month intervals, respectively. In addition, when patients complained of any symptoms or exhibited elevated tumor markers on blood tests, imaging studies were performed.
Protein expression by immunohistochemistry
The ERCC1 protein expression was evaluated by immunohistochemistry (IHC) using an anti-ERCC1 antibody (clone 8F1; Abcam, Cambridge, UK). We used a standard protocol for the immunostaining of the samples. The details of the method were previously described (35). Tumor nuclear staining intensity was graded on a scale of 0–3. The percentage of positive tumor nuclei was graded on a scale of 0–3. The percentage of positive tumor nuclei was evaluated, and a proportion score was attributed (0 if 0%; 0.1 if 1–9%; 0.5 if 10–49%; 1.0 if ≥ 50%), as previously described (36,37).
The antibody against TUBB3 was an anti-class III β-tubulin monoclonal antibody (clone TUJ1; Covance, Inc., Princeton, NJ, USA). Having over 50% of positive cells with a staining intensity of 2 was considered TUBB3-positive (35).
TS protein was evaluated by IHC using recombinant human TS-specific antibody (clone RTSSA; Taiho Pharmaceutical, Co., Ltd., Saitama, Japan). The slides were examined at low magnification, and the intensity of cytoplasmic staining was scored as follows: 0, no staining or faint staining; 1+, moderate staining; 2+, strong staining. We classified scores of 0 as negative and scores of 1+ and 2+ as positive for the TS antibody. We also judged cases with <10% of tumor cells with moderate or strong staining as being negative (38).
OPRT protein expression was evaluated by IHC using an anti-OPRT polyclonal antibody (Taiho Pharmaceutical, Co., Ltd.) The staining was the same as for TS (38). Scores of 0 and 1+ were classified as negative and scores of 2+ as positive for the OPRT protein. We also judged cases with <10% of tumor cells with moderate or strong staining as being negative.
DPD protein expression was evaluated by IHC using anti-DPD polyclonal antibody RDPDPA (dilution: 1:400; Taiho Pharmaceutical, Co., Ltd.) The staining was the same as the previously described method (28). Scores of 0 and 1+ were classified as negative and scores of 2+ as positive for the DPD protein. We also judged cases with <10% of tumor cells with moderate or strong staining as being negative.
All immunostained sections were evaluated by separate investigator without knowledge of the patients' clinical data to evaluated H-scoring accurately. Representative positive and negative cases of each IHC are shown in Fig. 1.
Statistical analysis
The sample size was determined based on a phase II study reported by Kawamura et al (39) applying docetaxel plus gemcitabine as an adjuvant chemotherapy in 35 patients. The number of patients in each arm was calculated using the Fleming method and found to be 32 per arm (32). However, sufficient data for patients in the study could not be gathered within the study period.
The characteristics, disease-free survival (DFS), and the overall survival (OS) of 38 patients who received over two courses of adjuvant chemotherapy were analyzed. The five-year DFS and OS were examined by the Kaplan-Meier method, and the difference in the two arms was calculated by the log-rank test. The differences in the rate of adverse events were evaluated by the χ2 test. All of the data were analyzed with the EZR software version 1.33 (www.jichi.ac.jp/saitama-sct/SaitamaHP.files/statmedEN.html) (40). P≤0.05 was considered to indicate a statistically significant difference.
ResultsPatients' characteristics
Forty patients with stage II or IIIA NSCLC who had received surgically complete resection were enrolled. Among the 40 patients, 2 were excluded due to the cessation of adjuvant chemotherapy because of a grade 4 allergic reaction (anaphylactic shock) induced by paclitaxel. The patients' characteristics are presented in Table I. Briefly, the patients were 7 females and 31 males ranging in age from 39–75 years, with a mean age of 63.6 years. There were no significant differences in the clinicopathological characteristics between arms A and B.
Protein expression on IHC
The ERCC1 IHC staining was positive in 18/38 cases (47%) in all patients. The positive cases were 10/19 (53%) in arm A and 8/19 (42%) in arm B, and there was no significant difference in the ERCC1 protein expression among the various adjuvant chemotherapy regimens. No association between the expression of ERCC1 and clinicopathological factors was identified (data not shown).
The TUBB3 IHC staining was positive in 17/38 cases (45%) in all patients. The positive cases were 9/19 (47%) in arm A and 8/19 (42%) in arm B, and there was no significant difference in the TUBB3 protein expression among adjuvant chemotherapy regimens. No association between the expression of TUBB3 and clinicopathological factors was identified (data not shown).
The TS IHC staining was positive in 21/38 cases (55%) in all patients. The positive cases were 11/19 (58%) in arm A and 10/19 (53%) in arm B, and there was no significant difference in the TS protein expression among adjuvant chemotherapy regimens. No association between the expression of TS and clinicopathological factors was identified (data not shown).
The OPRT IHC staining was positive in 16/38 cases (42%) in all patients. The positive cases were 7/19 (37%) in arm A and 9/19 (47%) in arm B, and there was no significant difference in the OPRT protein expression among adjuvant chemotherapy regimens. No association between the expression of OPRT and clinicopathological factors was identified (data not shown).
The DPD IHC staining was positive in 22/38 cases (58%) in all patients. The positive cases were 14/19 (74%) in arm A and 8/19 (42%) in arm B, and there was no significant difference in the DPD protein expression among adjuvant chemotherapy regimens. No association between the expression of DPD and clinicopathological factors was identified (data not shown).
The survival
The correlations between the OS plus DFS and the clinicopathological factors of the 38 patients are summarized in Table II. No factors, including the protein expression, were found to have significantly influenced the OS or DFS. Furthermore, there were no significant differences in the OS and DFS between the CP and S-1 adjuvant chemotherapy regimens. The 5-year OS and DFS of 38 patients was 81.0 and 59.6%, respectively (Fig. 2A and B). The Kaplan-Meier curves based on the adjuvant chemotherapy regimens are shown in Fig. 2C and D.
The correlations between the OS plus DFS and the clinicopathological factors of the 19 patients who received CP adjuvant chemotherapy are summarized in Table III. There were no factors found to have significantly influenced the OS or DFS in the patients who received the CP regimen. The protein expressions of ERCC1 and TUBB3 did not affect the OS or DFS.
The correlations between the OS plus DFS and the clinicopathological factors of the 19 patients who received S-1 adjuvant chemotherapy are summarized at Table IV. There were no factors found to have significantly influenced the OS in the patients who received the S-1 regimens. In the analysis of the DFS, the protein expression of TS was the only significant prognostic factor. However, the protein expression of TS did not affect the OS (Fig. 3A) or DFS (Fig. 3B) in the investigation of all 38 patients. Furthermore, the protein expression of TS did not affect the OS (Fig. 3C) in the investigation of the 19 patients who received S-1 adjuvant chemotherapy. However, when we limited our investigation to the DFS of the patients who received S-1 adjuvant chemotherapy, the TS-negative cases showed a longer DFS than the TS-positive cases (Fig. 3D).
Discussion
The survival of patients with advanced lung cancer is still unfavorable compared with malignant tumors of other organs (1). Recently, improved outcomes have been achieved with molecular-targeted therapy for select patients with epidermal growth factor receptor (EGFR)-activating mutations or ALK translocation (11–13). Understanding the genetic and molecular variations that affect the efficacy of chemotherapeutic agents may improve patient care by allowing physicians to optimize treatment for each patient. Even with cytotoxic anticancer drugs, it would be useful to know the factors predictive of a therapeutic effect before starting the administration of chemotherapy.
In this study, we evaluated the expression of several proteins in 38 patients with stage II and IIIA NSCLC who had received CP or S-1 as adjuvant chemotherapy. The 5-year OS and DFS of these 38 patients were 81.0 and 59.6%, respectively. These findings are comparable to those that have been reported recently (1,41). Concerning the OS analysis, the EGFR mutation status has been shown to influence the prognosis after relapse (11–13). Molecular-targeted therapeutic drugs apparently extend the OS in cases with EGFR mutations. It is therefore difficult to evaluate the effect of adjuvant chemotherapy on the OS in our small-scale study, because we don't have the data of gene mutations about all patients of this study. We should evaluate the DFS to clarify the relationship between protein expression and adjuvant chemotherapy efficacy. We should check the gene mutations (EGFR and ALK) to evaluated the effect of adjuvant chemotherapy on the OS in the future studies.
The CP regimen is considered as a standard chemotherapy regimen for recurrent and advanced lung cancer (42–46). We used the regimen of bi-weekly paclitaxel plus carboplatin to be able to complete the adjuvant chemotherapy without interruption due to side effects. As S-1 is considered more effective than UFT, long-term S-1 administration may be promising as an adjuvant chemotherapy regimen for advanced lung cancer (47). Indeed, several studies have shown that S-1 administration as adjuvant chemotherapy is associated with significant survival benefits following surgically complete resection for NSCLC (47,48). In this study, the 5-year OS and DFS were almost the same between the S-1 group and the CP group.
We investigated the protein expressions of ERCC1 and TUBB3, which are believed to be associated with the effect of platinum- and taxane-based chemotherapies, respectively. Previously, ERCC1-positive cases were reported to show more resistance to platinum-based chemotherapy than negative cases (16), but no relationship was noted between the ERCC1 expression and the prognosis, even in the patients who received the CP regimen in this study. We obtained similar findings concerning the TUBB3 expression. TUBB3-negative cases have previously been reported to show a better prognosis than positive ones. The prognostic effect of TUBB3 expression observed in this study, even in the patients who received CP regimen, was not consistent with prior published reports in the setting of advanced NSCLC (35,49,50). This discrepancy may be attributed to the small patient population in this study.
We also evaluated the protein expressions of TS, DPD, and OPRT, which are believed to be associated with the effect 5-FU-related agents, including S-1. Specifically, the overexpression of TS and DPD have been reported to be associated with resistance to S-1 (26–28). In contrast, the overexpression of OPRT was reported to be associated with a better prognosis in patients who received S-1 chemotherapy (30). In the present study, the expression of DPD and OPRT showed no association with the OS or DFS, even in the patients who received S-1 chemotherapy. The expression of TS did not have an association with the OS or DFS in the total population or with the OS in the 19 patients who received S-1. However, in the analysis of the DFS of the 19 patients who received S-1, the patients with TS overexpression showed a significantly poorer prognosis than the TS-negative patients.
One limitation associated with this study was the small patient population, as only 19 cases received S-1 and 19 cases received CP. Among the 40 patients, 2 were excluded due to the cessation of adjuvant chemotherapy because of a grade 4 allergic reaction (anaphylactic shock) induced by paclitaxel. The frequency of the anaphylactic shock (5%) was higher than previous reports. We think that the small sample size of this study will affect to the result. However, the adverse effects of S-1 were tolerable, and S-1 chemotherapy may be considered a promising adjuvant chemotherapy for patients with advanced disease who have undergone complete surgical resection. Further large-scale analyses of the relationship between TS expression and chemotherapeutic effects are desired. Moreover, we should evaluate the relationship among each protein expression in a large-scale clinical trial in the future.
We herein showed that TS is a potentially useful biomarker to help identify patients who will benefit from S-1 adjuvant chemotherapy.
Acknowledgements
Not applicable.
Funding
No funding was received.
Availability of data and material
The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.
Authors' contributions
KO and MY designed the present study. KO, KN, TY, TK, TN, MS, SM, HH, OK and MY collected the patients' data. KO, TT, TS, RO, TW and RN analyzed the patients' data. KO was a major contributor in writing the manuscript. All authors read and approved the final manuscript.
Ethics approval and consent to participate
Written informed consent was obtained from all patients, and the study protocol was approved by the Institutional Review Board of each participating institution (Nagoya City University Hospital no. 45-13-0020). This study was carried out in accordance with the Declaration of Helsinki.
Consent for publication
Written informed consent was obtained from all patients.
Competing interests
The authors declare that they have no competing interests.
ReferencesDetterbeckFCChanskyKGroomePBolejackVCrowleyJShemnskiLKennedyCKrasnikMPeakeMRami-PortaRThe IASLC lung cancer staging project: Methodology and validation used in the development of proposals for revision of the stage classification of NSCLC in the forthcoming (eighth) edition of the TNM classification of lung cancerJ Thorac Oncol1114331446201610.1016/j.jtho.2016.01.02427448762KisMGGasparLEChaftJEKennedyEBAzzoliCGEllisPMLinSHPassHISethRShepherdFAAdjuvant systemic therapy and adjuvant radiation therapy for stage I to IIIA completely resected non-small-cell lung cancers: American society of clinical oncology/cancer care ontario clinical practice guideline updateJ Clin Oncol3529602974201710.1200/JCO.2017.72.440128437162PistersKMEvansWKAzzoliCGKrisMGSmithCADeschCESomerfieldMRBrouwersMCDarlingGEllisPMCancer care onario and American society of clinical oncology adjuvant chemotherapy and adjuvant radiation therapy for stages I–IIIA resectable non-small cell lung cancer guidelineJ Clin Oncol2555065518200710.1200/JCO.2007.14.122617954710CrinòLWederWvan MeerbeeckJFelipEESMO Guidelines Working Group: Early stage and locally acvanced (non-metastatic) non-small-cell lung cancer: ESMO clinical practice guidelines for diagnosis, treatment and follow-upAnn Oncol21Suppl 5v103v115201010.1093/annonc/mdq20720555058NSCLC Meta-analyses Collaborative GroupArriagadaRAuperinABurdettSHigginsJPJohnsonDHLe ChevalierTLe PechouxCParmarMKPignonJPAdjuvant chemotherapy, with or without postoperative radiotherapy, in operable non-small-cell lung cancer: Two meta-analyses of individual patient dataLancet37512671277201010.1016/S0140-6736(10)60059-120338627FrühMRollandEPignonJPSeymourLDingKTribodetHWintonTLe ChevalierTScagliottiGVDouillardJYPooled analysis of the effect of age on adjuvant cisplatin-based chemotherapy for completely resected non-small-cell lung cancerJ Clin Oncol2635733581200810.1200/JCO.2008.16.272718640938HottaKMatsuoKUeokaHKiuraKTabataMTanimotoMRole of adjuvant chemotherapy in patients with resected non-small-cell lung cancer: Reappraisal with meta-analysis of randomized controlled trialsJ Clin Oncol2238603867200410.1200/JCO.2004.02.10915326194SedrakyanAVan Der MeullenJO'ByrneKPrendivilleJHillJTreasureTPostoperative chemotherapy for non-small cell lung cancer: A systematic review and meta-analysisJ Thorac Cardiovasc Surg128414419200410.1016/j.jtcvs.2004.04.02715354101BerghmansTPaesmansMMeertAPMascauxCLothairePLafitteJJSculierJPSurvival improvement in resectable non-small cell lung cancer with (neo)adjuvant chemotherapy: Results of a meta-analysis of the literatureLung Cancer491323200510.1016/j.lungcan.2005.01.00215949586KellyKCrowleyJBunnPAJrPresantCAGrevstadPKMoinpourCMRamseySDWozniakAJWeissGRMooreDFRandomized phase III trial of paclitaxel plus carboplatin versus vinorelbine plus cisplatin in the treatment of patients with advanced non-small-cell lung cancer: A Southwest Oncology Group trialJ Clin Oncol1932103218200110.1200/JCO.2001.19.13.321011432888LynchTJBellDWSordellaRGurubhagavatulaSOkimotoRABranniganBWHarrisPLHaserlatSMSupkoJGHaluskaFGActivating mutations in the epedermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinibN Engl J Med35021292139200410.1056/NEJMoa04093815118073PaezJGJännePALeeJCTracySGreulichHGabrielSHermanPKayeFJLindemanNBoggonTJEGFR mutations in lung cancer: Correlation with clinical response to gefitinib therapyScience30414971500200410.1126/science.109931415118125KwakELBangYJCamidgeDRShawATSolomonBMakiRGOuSHDezubeBJJännePACostaDBAnaplastic lymphoma kinase inhibition in non-small-cell lung cancerN Engl J Med36316931703201010.1056/NEJMoa100644820979469WangDLippardSJCellular processing of platinum anticancer drugsNature Rev Drug Disc4307320200510.1038/nrd1691AltahaRLiangXYuJJReedEExcision repair cross complementing-group 1: Gene expression and platinum resistanceInt J Mol Med14959970200415547660LordRVBrabenderJGandaraDAlberolaVCampsCDomineMCardenalFSánchezJMGumerlockPHTarónMLow ERCC1 expression correlates with prolonged survival after cisplatin plus gemcitabine chemotherapy in non-small cell lung cancerClin Cancer Res822862291200212114432MetzgerRLeichmanCGDanenbergKDDanenbergPVLenzHJHayashiKGroshenSSalongaDCohenHLaineLERCC1 mRNA levels complement thymidylate synthase mRNA levels in predicting response and survival for gastric cancer patients receiving combination cisplatin and fluorouracil chemotherapyJ Clin Onclo16309316199810.1200/JCO.1998.16.1.309BurkhartCAKavallarisMHorwitzBand SThe role of beta-tubulin isotypes in resistance to antimitotic drugsBiochem Biophys Acta1471O19200111342188KatsetosCDLegidoAPerentesEMörkSJClass III beta-tubulin isotype: a key cytoskeletal protein at the crossroads of developmental neurobiology and tumor neuropathologyJ Child Neurol18851867200310.1177/08830738030180120514736079LuQLuduenaRFRemoval of beta III isotype enhances taxol induced microtubule assemblyCell Struct Funct18173182199310.1247/csf.18.1737902216KamathKWilsonLCabralFJordanMABetaIII-tubulin induces paclitaxel resistance in association with reduced effects on microtubule dynamic instabilityJ Biol Chem2801290212907200510.1074/jbc.M41447720015695826LenzHJLeichmanCGDanenbergKDDanenbergPVGroshenSCohenHLaineLCrookesPSilbermanHBarandaJThymidylate synthase mRNA level in adenocarcinoma of the stomach: A predictor for primary tumor response and overall survivalJ Clin Oncol14176182199610.1200/JCO.1996.14.1.1768558194TakedaMOkamotoIHirabayashiNKitanoMNakagawaKThymidylate synthase and dihydropyrimidine dehydrogenase expression levels are associated with response to S-1 plus carboplatin in advanced non-small cell lung cancerLung Cancer73103109201110.1016/j.lungcan.2010.10.02221111509OgiuchiYMaruokaYAndoTKobayashiMOgiuchiHThymidylate synthase, thymidine phosphorylase and orotate phosphoribosyl transferase levels as predictive factors of chemotherapy in oral squzmous cell carcinomaActa Histochem Cytochem413946200810.1267/ahc.0800218636111DiasioRBHarrisBEClinical pharmacology of 5-fluorouracilClin Parmacokinet16215237198910.2165/00003088-198916040-00002OguriTAchiwaHBesshoYMuramatsuHMaedaHNiimiTSatoSUedaRThe role of thymidylate synthase and dihydropyrimidine dehydrogenase in resistance to 5-fluorouracil in human lung cancer cellsLung Cancer49345351200510.1016/j.lungcan.2005.05.00315993511EtienneMCChéradameSFischelJLFormentoPDassonvilleORenéeNSchneiderMThyssADemardFMilanoGResponse to fluorouracil therapy in cancer patients: The role of tumoral dihydorpyrimidine dehydrogenase activityJ Clin Oncol1316631670199510.1200/JCO.1995.13.7.16637602356MiyoshiTKondoKTobaHYoshidaMFujinoHKenzakiKSakiyamaSTakehisaMTangokuAPredictive value of thymidylate synthase and dihydropyrimidine dehydrogenase expression in tumor tissue, regarding the efficacy of postoperatively administered UFT (Tegafur+Uracil) in patients with non-small cell lung cancerAnticancer Res2726412648200717695427SakuraiYSakamotoKSugimotoYYoshidaIMasuiTTonomuraSInabaKShojiMNakamuraYUyamaIOrotate phosphoribosyltransferase levels measured by a newly established enzyme-linked immunosorbent assay in gastric carcinomaCancer Sci97492498200610.1111/j.1349-7006.2006.00200.x16734727ChoiISLeeHSLeeKWKimHKimKHKimYJKimJHKimWHLeeJSBiomarker analysis in patients with advanced gastric cancer treated with S-1 plus cisplatin chemotherapy: Orotate phosphoribosyltransferase expression is associated with treatment outcomesMed Oncol28991998201110.1007/s12032-010-9590-420533001SobinLHGospodarowiczMKWittekindCHTNM Classification of Malignant Tumours7th editionWiley-BlackwellOxford2009OkudaKYanoMTatematsuTNakamaeKYamadaTKasugaiTNishidaTSanoMMoriyamaSHanedaHS-1 vs. paclitaxel plus carboplatin as adjuvant chemotherapy for completely resected stage II/IIIA non-small-cell lung cancerMol Clin Oncol87379201829387399CalvertAHNewellDRGumbrellLAO'ReillySBurnellMBoxallFESiddikZHJudsonIRGoreMEWiltshawECarboplatin dosage: Prospective evaluation of a simple formula based on renal functionJ Clin Oncol717481756198910.1200/JCO.1989.7.11.17482681557JellifieRWJlliffeSMA computer program for estimation of creatine clearance from unstable serum creatine levels, age, sex, and weightMath Biosci141724197210.1016/0025-5564(72)90003-XOkudaKSasakiHDumontetCKawanoOYukiueHYokoyamaTYanoMFujiiYExpression of excision repair cross-complementation group 1 and class III beta-tubulin predict survival after chemotherapy for completely resected non-small cell lung cancerLung Cancer62105112200810.1016/j.lungcan.2008.02.02118395930Al HaddadSZhangZLeygueESnellLHuangANiuYHiller-HitchcockTHoleKMurphyLCWatsonPHPsoriasin (S100A7) expression and invasive breast cancerAm J Pathol15520572067199910.1016/S0002-9440(10)65524-110595935Handara-LucaABilalHBertrandLCFouretPExtra-cellular signal-regulated ERK-1/ERK-2 pathway activation in human salivary gland mucoepidermoid carcinoma: association to aggressive tumor behavior and tumor cell proliferationAm J Pathol163957967200310.1016/S0002-9440(10)63455-412937136YokotaKSasakiHOkudaKShitaraMHikosakaYMoriyamaSYanoMFujiiYExpression of thymidylate synthase and orotate phosphoribosyltransferase in thymic carcinomaExp Ther Med4589593201210.3892/etm.2012.64123170110KawamuraMEguchiKIzumiYYamatoYKoikeTSakaguchiHHadaEKobayashiKPhase II trial of gemcitabine and docetaxel in patients with completely resected stage IIA-IIIA non-small-cell lung cancerCancer Chemother Pharmacol60495501200710.1007/s00280-006-0391-617143600KandaYInvestigation of the freely available easy-to-use sofrware ‘EZR’ for medical statisticsBone Marrow Transplant48452458201310.1038/bmt.2012.24423208313AsamuraHGoyaTKoshiishiYSoharaYEguchiKMoriMNakanishiYTsuchiyaRShimokataKInoueHA Japanese lung cancer registry study: Prognosis of 13,010 resected lung cancersJ Thorac Oncol34652200810.1097/JTO.0b013e31815e857718166840OheYOhashiYKubotaKTamuraTNakagawaKNegoroSNishiwakiYSaijoNAriyoshiYFukuokaMRandomized phase III study of cisplatin plus irinotecan versus carboplatin plus paclitaxel, cisplatin plus gemcitabine, and cisplatin plus vinorelbine for advanced non-small-cell lung cancer: Four-Arm cooperative study in JapanAnn Oncol18317323200710.1093/annonc/mdl37717079694SchillerJHHarringtonDBelaniCPLangerCSandlerAKrookJZhuJJohnsonDHEastern Cooperative Oncology Group: Comparison of four chemotherapy regimens for advanced non-small-cell lung cancerN Engl J Med3469298200210.1056/NEJMoa01195411784875YamashitaYKataokaKIshidaTMatsuuraMSenoNMukaidaHMiyaharaEMiyataYOkitaRShimizuKA feasibility study of postoperative adjuvant therapy of carboplatin and weekly paclitaxel for completely resected non-small cell lung cancerJ Thorac Oncolo3612616200810.1097/JTO.0b013e318174e05eIchikiMKawasakiMTakayamaKNinomiyaKKubaMIwamiFMiyazakiNOishiKTakeoSAizawaHNakanishiYA multi-center phase II study of carboplatin and paclitaxel with a biweekly schedule in patients with advanced non-small-cell lung cancer. Kyushu thoracic oncology group trialCancer Chemother Pharmacol58368373200610.1007/s00280-005-0168-316395589SugayaMUramotoHUchiyamaANagashimaANakanishiRSakataHNakanishiKHanagiriTYasumotoKPhase II trial of adjuvant chemotherapy with bi-weekly carboplatin plus paclitaxel in patients with completely resected non-small cell lung cancerAnticancer Res3030393044201020683052IwamotoYMitsudomiTSakaiKYamanakaTYoshiokaHTakahamaMYoshimuraMYoshinoITakedaMSugawaraSRandomized phase II study of adjuvant chemotherapy with long-term S-1 versus Cisplatin+S-1 in completely resected stage II–IIIA non-small cell lung cancerClin Cancer Res2152455252201510.1158/1078-0432.CCR-14-316026253869OkumuraSSasakiTSatohKKitadaMNagaseAYatsuyanagiEOhsakiYFeasibility of adjuvant chemotherapy with S-1 consisting of a 4-week administration and a two-week rest period in patients with completely resected non-small cell lung cancerMol Clin Oncol1124130201310.3892/mco.2012.624649134MozzettiSFerliniCConcolinoPFilippettiFRaspaglioGPrisleiSGalloDMartinelliERanellettiFOFerrandinaGScambiaGCClass III beta-tubulin overexpression is a prominent mechanism of paclitaxel resistance in ovarian cancer patientsClin Cancer Res11298305200515671559ParadisoAMangiaAChiriattiATommasiSZitoALatorreASchttulliFLorussoVBiomarkers predictive for clinical efficacy of taxol-based chemotherapy in advanced breast cancerAnn Oncol16Suppl 4iv14iv19200510.1093/annonc/mdi90215923415
Kaplan-Meier analyses. (A) The 5-year overall survival of the 38 patients who received carboplatin plus paclitaxel or S-1 adjuvant chemotherapy. (B) The 5-year disease-free survival of the 38 patients who received carboplatin plus paclitaxel or S-1 adjuvant chemotherapy. (C) The overall survival divided by carboplatin plus paclitaxel or S-1 adjuvant chemotherapy. (D) The disease-free survival divided by carboplatin plus paclitaxel or S-1 adjuvant chemotherapy.
Kaplan-Meier analyses. (A) The overall survival of the 38 patients associated withthe protein expression of TS. (B) The disease-free survival of the 38 patients associated with the protein expression of TS. (C) The overall survival of the 19 patients who received S-1 adjuvant chemotherapy, associated with the protein expression of TS. (D) The disease-free survival of the 19 patients who received S-1 adjuvant chemotherapy, associated with the protein expression of TS. TS, thymidylate synthase.
Characteristics of the 38 patients recruited to the present study.
Characteristics
All patients (n=38)
CBDCA+PTX (n=19)
S-1 (n=19)
P-value
Observation period, months
15–98/67
19–98/67
15–87/67
0.951
Sex, n
Male
31
14
17
0.405
Female
7
5
2
Age, years
39–75/63.6
47–73/64.4
39–75/62.9
0.529
Histological type, n
Adenocarcinoma
24
11
13
0.737
Squamous cell carcinoma
13
7
6
Others
1
1
0
Pathological stage (IIA/IIB/IIIA), n
17/11/10
9/5/5
8/6/5
0.980
ERCC1 (Positive/negative), n
18/20
10/9
8/11
0.746
TUBB3 (Positive/negative), n
17/21
9/10
8/11
0.980
TS (Positive/negative), n
21/17
11/8
10/9
0.980
OPRT (Positive/negative), n
16/22
7/12
9/10
0.743
DPD (Positive/negative), n
22/16
14/5
8/11
0.091
Data are presented as the range/median, or as the n number of patients. PTX, paclitaxel; CBDCA, carboplatin; ERCC1, excision repair cross-complementation group 1; TUBB3, class III β-tubulin; TS, thymidylate synthase; OPRT, orotate phosphoribosyltransferase; DPD, dihydropyrimidine dehydrogenase.
Correlation with overall survival plus disease free survival and clinicopathological factors.
Overall survival
Disease free survival
Factor
Subgroup
Total n (n=38)
5-year survival (%)
P-value
5-year survival (%)
P-value
Age, years
≤65/>65
20/18
78.8/83.3
0.182
63.6/55.6
0.898
Sex
Male/female
31/7
76.5/100
0.070
56.8/71.4
0.398
Tissue type
Adenocarcinoma/others
24/14
87.1/70.0
0.399
58.3/60.2
0.477
Pathological stage
IIA/IIB or IIIA
17/21
87.8/75.2
0.085
69.1/52.4
0.250
Chemotherapy regime
CP/S-1
19/19
78.6/83.6
0.976
50.8/68.4
0.351
ERCC1
Positive/negative
18/20
76.0/85.0
0.773
70.6/50.0
0.111
TUBB3
Positive/negative
17/21
87.5/75.6
0.696
64.7/54.8
0.869
TS
Positive/negative
21/17
74.4/88.2
0.092
50.1/70.6
0.140
OPRT
Positive/negative
16/22
86.2/77.3
0.783
66.1/54.5
0.502
DPD
Positive/negative
22/16
86.1/73.7
0.824
66.5/50.0
0.331
CP, carboplatin plus paclitaxel; ERCC1, excision repair cross-complementation group 1; TUBB3, class III β-tubulin; TS, thymidylate synthase; OPRT, orotate phosphoribosyltransferase; DPD, dihydropyrimidine dehydrogenase.
Correlation with overall survival plus disease free survival and clinicopathological factors for Carboplatin plus paclitaxel patients.
Overall survival
Disease free survival
Factor
Subgroup
Total n (n=19)
5-year survival (%)
P-value
5-year survival (%)
P-value
Age, years
≤65/>65
9/10
76.2/80.0
0.598
63.5/40.0
0.281
Sex
Male/female
14/5
70.7/100
0.134
47.6/60.0
0.601
Tissue type
Adenocarcinoma/others
11/8
90.9/60.0
0.325
36.4/72.9
0.172
Pathological stage
IIA/IIB or IIIA
9/10
76.2/80.0
0.473
53.3/50.0
0.902
ERCC1
Positive/negative
10/9
58.3/100
0.773
67.5/33.3
0.129
TUBB3
Positive/negative
9/10
77.8/78.7
0.527
44.4/57.1
0.502
TS
Positive/negative
11/8
71.6/87.5
0.310
53.0/50.0
0.700
OPRT
Positive/negative
7/12
68.6/83.3
0.824
68.6/41.7
0.321
DPD
Positive/negative
14/5
77.9/80.0
0.806
62.9/20.0
0.164
CP, carboplatin plus paclitaxel; ERCC1, excision repair cross-complementation group 1; TUBB3, class III β-tubulin; TS, thymidylate synthase; OPRT, orotate phosphoribosyltransferase; DPD, dihydropyrimidine dehydrogenase.
Correlation with overall survival plus disease free survival and clinicopathological factors for S-1 patients.
Overall survival
Disease free survival
Factor
Subgroup
Total n (n=19)
5-year survival (%)
P-value
5-year survival (%)
P-value
Age, years
≤65/>65
11/8
80.8/87.5
0.202
63.6/75.0
0.389
Sex
Male/female
17/2
81.6/100
0.316
64.7/100
0.297
Tissue type
Adenocarcinoma/others
13/6
83.9/83.3
0.839
76.9/50.0
0.570
Pathological stage
IIA/IIB or IIIA
8/11
100/70.7
0.065
87.5/54.5
0.072
ERCC1
Positive/negative
8/11
100/72.7
0.142
75.0/63.6
0.333
TUBB3
Positive/negative
8/11
100/72.7
0.241
87.5/54.5
0.346
TS
Positive/negative
10/9
77.8/88.9
0.187
50.0/88.9
0.044
OPRT
Positive/negative
9/10
100/70.0
0.496
66.7/70.0
0.783
DPD
Positive/negative
8/11
100/70.7
0.587
75.0/63.6
0.721
CP, carboplatin plus paclitaxel; ERCC1, excision repair cross-complementation group 1; TUBB3, class III β-tubulin; TS, thymidylate synthase; OPRT, orotate phosphoribosyltransferase; DPD, dihydropyrimidine dehydrogenase.