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EGFR signaling has been implicated in hypoxia-associated resistance to radiation or chemotherapy. Non–small cell lung carcinomas (NSCLC) with activating L858R or ΔE746-E750 EGFR mutations exhibit elevated EGFR activity and downstream signaling. Here, relative to wild-type (WT) EGFR, mutant (MT) EGFR expression significantly increases radiosensitivity in hypoxic cells. Gene expression profiling in human bronchial epithelial cells (HBEC) revealed that MT-EGFR expression elevated transcripts related to cell cycle and replication in aerobic and hypoxic conditions and downregulated RAD50, a critical component of nonhomologous end joining and homologous recombination DNA repair pathways. NSCLCs and HBEC with MT-EGFR revealed elevated basal and hypoxia-induced γ-H2AX–associated DNA lesions that were coincident with replication protein A in the S-phase nuclei. DNA fiber analysis showed that, relative to WT-EGFR, MT-EGFR NSCLCs harbored significantly higher levels of stalled replication forks and decreased fork velocities in aerobic and hypoxic conditions. EGFR blockade by cetuximab significantly increased radiosensitivity in hypoxic cells, recapitulating MT-EGFR expression and closely resembling synthetic lethality of PARP inhibition. Implications: This study demonstrates that within an altered DNA damage response of hypoxic NSCLC cells, mutant EGFR expression, or EGFR blockade by cetuximab exerts a synthetic lethality effect and significantly compromises radiation resistance in hypoxic tumor cells. Mol Cancer Res; 15(11); 1503–16. ©2017 AACR.

Background: Chronic obstructive pulmonary disease is a smoking-related disease, and is categorized into the emphysema and airway dominant phenotypes. We examined the relationship between emphysematous changes and epidermal growth factor receptor (EGFR) mutation status in patients with lung adenocarcinoma. Patients and Methods: The medical records for 250 patients with lung adenocarcinoma were retrospectively reviewed. All patients were categorized into the emphysema or non-emphysema group. Results: Wild-type EGFR was detected in 136 (54%) and mutant EGFR in 48 (19%). Emphysematous changes were observed in 87 (36%) patients. EGFR mutation was highly frequent in the non-emphysema group (p=0.0014). Multivariate logistic regression analysis showed that emphysema was an independent risk factor for reduced frequency of EGFR mutation (Odds Ratio=3.47, p=0.005). Conclusion: Our data showed a relationship between emphysematous changes and EGFR mutation status. There might be mutually exclusive genetic risk factors for carcinogenesis and development of emphysematous changes.

Immune checkpoint inhibitors (ICIs), including nivolumab, have exhibited substantial benefits in the treatment of several types of cancers. However, treatment with ICIs is often accompanied by immune-related adverse events (irAEs), and a clear understanding of the precise indications and management of irAEs is important for harnessing the full potential of these agents. While skin- or gastrointestinal-associated irAEs have been relatively well studied, there are few reports regarding nivolumab-induced cholangitis. We retrospectively reviewed data from patients with advanced or recurrent non-small cell lung cancer who were treated with nivolumab between December 2015 and December 2018 at Tottori University in Japan. Among the 59 patients, we identified four patients who experienced nivolumab-induced cholangitis. Of these four patients, stable disease (SD) was observed in two patients (50%), while partial response (PR) was achieved in two patients (50%) under nivolumab treatment. Patients were treated with corticosteroid alone (n=2) or in combination with mycophenolate mofetil (MMF) (n=2); these treatments resulted in improvements in nivolumab-induced cholangitis in three patients. In conclusion, the present retrospective study identified four cases of nivolumab-induced cholangitis. The combination of corticosteroid and MMF was effective in two cases with grade 4 nivolumab-induced cholangitis. Further reports are needed to establish the optimal management of patients with this irAE.

Background Precision medicine based on driver oncogenes is now developed for non-small cell lung cancer. Recent approval of next-generation sequencing (NGS)-based companion diagnostics has increased the need for the use of high-quality specimens for the diagnosis. Methods From August 2015 to May 2018, 82 patients who were enrolled in a nation-wide genome screening, LC-SCRUM-Japan, from our institution were examined for the success rates of genetic analysis according to sample types. A NGS analysis, Oncomine Comprehensive Assay, was performed using DNA and RNA extracted from lung cancer specimens. Results The success rates of DNA and RNA analyses were 100% (82/82 cases) and 78% (64/82), respectively. Of the 82 samples analyzed, 80 (98%) were tissues and two (2%) were pleural effusions. The success rates of RNA analysis in the 80 tissue samples were 45/60 (73%) in bronchoscopic byopsy, 10/10(100%) in operation, 5/6(83%) in Percutaneous biopsy and 3/4(75%) in others. Of the 60 bronchoscopic samples, the success rates were 72% (18/25) in EBUS-TBNA, 88% (7/8) in EBUS-GS (large diameter), 69% (11/16) in EBUS-GS (small diameter) and 82% (9/11) in direct vision biopsy. In the 82 cases analyzed, a total of 52 actionable gene alterations (18 KRAS mut, 6 EGFR mut, 6 MET amp/fus, 5 RET fus, 5 ALK fus, 4 PIK3CA mut, 2 ERBB2 mut, 2 ROS1 fus, 2 FGFR1 amp, 1 BRAF mut, 1 FGFR3 fus) were detected in 48 cases (59%). Twelve of the 48 (15%) were registered in ongoing clinical trials of targeted drugs. Conclusions All the samples were available for DNA analysis of the NGS. The success rates of RNA analysis were lower in samples obtained from EBUS-TBNA and EBUS-GS (small diameter), suggesting that the success rates depend on the sample size.

Objectives Zoledronate (ZOL) is usually used for prevention of skeletal-related events in cancer patients with bone metastases. The first administration of ZOL is occasionally associated with development of acute-phase reaction (APR), which is due to activation of γδ T cells. ZOL-related APR was associated with better overall survival (OS) of patients with non-small cell lung cancer (NSCLC) in our previous retrospective study. However, it remains to be clarified whether γδ T cells are more activated in patients who experienced ZOL-related APR, and whether γδ T cell activation is involved in prolongation of OS. Materials and Methods Twenty-three patients with advanced NSCLC were recruited between 2012 and 2014 in this study. We administered ZOL to participants with standard care. The patient characteristics, change in γδ T cell counts and cytokines, OS, and skeletal-related event-free survival were compared between patients with APR (APR group) and those without APR (non-APR group). Results Ten patients (43.5%) experienced a ZOL-related APR. The number of γδ T cells at baseline in the APR group was significantly higher than that in the non-APR group. Serum interleukin-6 and tumor necrosis factor-α in the APR group were significantly increased, but no change in the number of γδ T cells was observed after the first administration of ZOL in both groups. OS in the APR group was significantly longer than that in the non-APR group (median survival time: 23.1 vs. 14.5 months, p Conclusion We showed that APR is related to higher numbers of γδ T cells at baseline and increased cytokines after the first ZOL administration, but not to proliferative responses of γδ T cells. In addition, better OS was observed in the APR group. Therefore, the number of γδ T cells might be a prognostic marker in patients with NSCLC.

Epidermal growth factor receptor (EGFR) gene mutation testing is essential for choosing appropriate treatment options in patients with advanced non-small cell lung cancer (NSCLC). However, a time delay occurs between histological diagnosis and molecular diagnosis in clinical situations. To minimize this delay, we developed a novel point-of-care test for EGFR mutations, based on a high-speed real-time polymerase chain reaction (PCR) system designated here as ultrarapid PCR combined with highly accurate bronchoscopic sampling. We investigated whether our system for detecting EGFR mutations was valid by comparing test results with those obtained using a commercialized EGFR mutation test. We obtained small amounts of bronchial lavage fluids after transbronchial biopsies (TBBs) were performed on enrolled patients (n=168) who underwent endobronchial ultrasonography using a guide sheath (EBUS-GS). EGFR mutation analysis was performed by ultrarapid PCR immediately after EBUS-GS-TBBs were obtained (on the same day). After pathological diagnoses of NSCLC, EGFR mutation status in formalin-fixed, paraffin- embedded samples was confirmed by the PCR-invader method, and the concordance rates between the PCR methods were compared. The total diagnostic yield of EBUS-GS-TBB was 91.0%. The positive concordance rates for detecting 19del and L858R with the ultrarapid PCR and PCR-invader methods were both 100%. Negative concordance rates were 97.2 and 98.1%, respectively. We also demonstrated a dramatic effect of early erlotinib administration, based on ultrarapid PCR results, for a 52-year-old woman suffering from respiratory failure due to severe intrapulmonary metastases with poor performance status. In conclusion, ultrarapid PCR combined with EBUS-GS-TBB enabled rapid and reliable point-of-care testing for EGFR mutations.

Aromatase inhibitors are superior to tamoxifen as adjuvant therapy in postmenopausal patients with hormone-responsive breast cancer. We report the follow-up efficacy results from the N-SAS BC 03 trial (UMIN CTRID: C000000056) where anastrozole was compared with tamoxifen as adjuvant therapy in postmenopausal Japanese patients with hormone-responsive early breast cancer. The full analysis set contained 696 patients (anastrozole arm, n = 345; tamoxifen arm, n = 351). The log-rank test was used to compare the two groups in terms of disease-free survival (DFS) and relapse-free survival (RFS); Kaplan–Meier estimates were calculated. The treatment effects were estimated by Cox’s proportional hazards model. To examine time-varying effect of hazard ratios, we estimated time-varying hazard ratios at time t [HR(t)] using data from time t up to 12 months. After a median follow-up of 98.5 months, hazard ratios (95 % CIs) were 0.90 (0.65–1.24; log-rank p = 0.526) for DFS and 0.83 (0.56–1.23; log-rank p = 0.344) for RFS. Hazard ratios (95 % CIs) for DFS and RFS up to 36 months were 0.69 (0.40–1.17) and 0.54 (0.27–1.06) and those after 36 months were 1.06 (0.70–1.59) and 1.05 (0.64–1.73), respectively. Time-varying hazard ratios for both DFS and RFS showed that hazard ratios were initially in favor of anastrozole and approached 1.0 at around 36 months. Superior efficacy of anastrozole to tamoxifen suggested by the initial analysis was not confirmed in the present analysis after a long-term follow-up period. Advantage of anastrozole was the greatest immediately after switching from tamoxifen and then decreased thereafter.

Previously we showed that Akt-suppressing agents, combined with amrubicin, synergistically inhibited the growth of small cell lung cancer cells. The combined effects of chemotherapeutic agents and Akt-suppressing agents, including epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, were evaluated in A549 lung adenocarcinoma cells harboring K-ras mutation and wild-type EGFR. Only amrubicin and not other chemotherapeutics (cisplatin, pemetrexed and paclitaxel) synergistically inhibited cell growth when combined with an Akt inhibitor, LY294002. The combination of amrubicin and LY294002 enhanced Annexin V binding to cells. A non-specific tyrosine kinase inhibitor, genistein, suppressed Akt and showed synergistic interaction in combination with amrubicin. Two EGFR tyrosine kinase inhibitors (EGFR-TKIs), gefitinib and erlotinib, suppressed Akt activity at clinically achievable concentrations and demonstrated synergism when combined with amrubicin. The suppression of K-ras expression by siRNA interfered with this synergism and inhibited both EGFR and Akt activity in A549 cells. In Ma10 cells, which harbor wild-type EGFR and K-ras, EGFR-TKIs neither suppressed Akt activity nor exhibited such synergism when combined with amrubicin. We concluded that the synergism by the combination of EGFR-TKI and amrubicin is attributable, at least partially, to K-ras mutation in A549 cells. The combination of EGFR-TKI and amrubicin may be a promising treatment for lung cancer with wild-type EGFR and K-ras mutation.

The EGF receptor (EGFR) contributes to tumor radioresistance, in part, through interactions with the catalytic subunit of DNA-dependent protein kinase (DNA-PKc), a key enzyme in the nonhomologous end joining DNA repair pathway. We previously showed that EGFR-DNA-PKcs interactions are significantly compromised in the context of activating mutations in EGFR in non–small cell lung carcinoma (NSCLC) and human bronchial epithelial cells. Here, we investigate the reciprocal relationship between phosphorylation status of DNA-PKcs and EGFR-mediated radiation response. The data reveal that both the kinase activity of DNA-PKcs and radiation-induced phosphorylation of DNA-PKcs by the ataxia telangiectasia–mutated (ATM) kinase are critical prerequisites for EGFR-mediated radioresponse. Alanine substitutions at seven key serine/threonine residues in DNA-PKcs or inhibition of DNA-PKcs by NU7441 completely abrogated EGFR-mediated radioresponse and blocked EGFR binding. ATM deficiency or ATM inhibition with KU55933 produced a similar effect. Importantly, alanine substitution at an ATM-dependent DNA-PKcs phosphorylation site, T2609, was sufficient to block binding or radioresponse of EGFR. However, mutation of a DNA-PKcs autophosphorylation site, S2056 had no such effect indicating that DNA-PKcs autophosphorylation is not necessary for EGFR-mediated radioresponse. Our data reveal that in both NSCLCs and human bronchial epithelial cells, activating mutations in EGFR specifically abolished the DNA-PKcs phosphorylation at T2609, but not S2056. Our study underscores the critical importance of a reciprocal relationship between DNA-PKcs phosphorylation and EGFR-mediated radiation response and elucidates mechanisms underlying mutant EGFR-associated radiosensitivity in NSCLCs. Mol Cancer Res; 10(10); 1359–68. ©2012 AACR.

The aim of the study was to determine the feasibility and validity of a newly developed patient-based instrument--the Patient Neurotoxicity Questionnaire (PNQ)--for grading chemotherapy-induced peripheral neuropathy (CIPN).We prospectively collected data from 300 female patients who were treated with taxane chemotherapy for primary breast cancer as part of a national multicenter phase III randomized trial (N-SAS BC 02). We evaluated patient compliance with the PNQ and several validation parameters, including concordance between CIPN grades noted by physicians (National Cancer Institute Common Toxicity Criteria) and patients (PNQ), and the concurrent validity and responsiveness of the PNQ versus the Functional Assessment of Cancer Therapy/Gynecologic Oncology Group-Neurotoxicity (FACT/GOG-Ntx) utilizing data at pre-treatment and before three, five, and seven treatment cycles.The questionnaire completion rate was90% at all assessments. Evaluation by physicians always resulted in lower neuropathy assessment scores compared with those reported directly by patients (weighted kappa coefficients, 0.02-0.06). Both PNQ sensory and motor scores were significantly correlated with the FACT/GOG-Ntx (r = 0.66 and 0.51, respectively). In the repeated measures analysis of variance model, PNQ grades increased considerably as treatment continued, indicating progressively worsening CIPN over time.The PNQ has an applicable degree of feasibility and validity, useful for the diagnosis of CIPN as well as for clinical treatment decision-making, where the development of CIPN is a potential treatment-limiting consideration. Physicians underreport and underestimate the severity of CIPN symptoms compared with patients, thereby supporting the importance of assessing patient-reported outcomes using the PNQ.

FDG-PETが診断に有用であった直腸癌甲状腺転移を経験したので報告する. 症例は60歳の女性で, 平成13年9月に直腸癌に対し腹会陰式直腸切断術を施行した. 平成14年1月に肺転移に対し左肺楔状切除を施行した. 術後l-LV・5-FU療法を7コース施行したが, 次第にCEAの上昇を認めたためCTおよびシンチグラムなどで全身検索を行ったが明らかな転移巣および局所再発を指摘できなかった. 12月にFDG-PETを施行したところ左頸部にFDGの集積が指摘された. 直腸癌甲状腺転移の診断で平成16年1月甲状腺左葉切除を施行した. 病理組織学的検査所見は腺癌の甲状腺転移であった. 直腸癌の甲状腺転移は非常にまれであり, 本邦報告例は自験例を含め5例であった. FDG-PETは, 従来の画像検査で異常を指摘できないが腫瘍マーカーの上昇を来している直腸癌術後患者の転移の検索に有用であると考えられた.

乳癌の消化管転移,なかでも大腸への転移は臨床では極めて稀である.われわれは乳腺浸潤性小葉癌術後で大腸転移をきたした1例を経験したので報告する.症例は45歳,女性.腹痛,嘔吐を主訴に来院し,腸閉塞と診断された.消化管造影,大腸内視鏡検査で下行結腸に完全狭窄が認められ,生検では悪性所見は認められなかったが,原発性または転移性大腸癌を考え,手術を施行した.下行結腸以外にも複数の腫瘍があり,さらにリンパ節転移や播種性病変も認められ,根治性なしと判断した.腸閉塞解除目的に腸管切除を施行し,病理組織学的検査で乳癌の転移と診断された.ゴセレリン・タモキシフェン併用療法を施行し,術後9カ月経過しSDを継続中である.乳癌患者において腹部症状の訴えの際には消化管転移の可能性を考える必要がある.また,消化管転移をきたした状態でもホルモン療法,化学療法の効果が期待できるため手術でのQOLの改善は有用である.

Although cytotoxic chemotherapy is essential in epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer (NSCLC), it is unclear which regimen is most effective. We retrospectively compared the efficacy of standard platinum-based chemotherapy with that of combination chemotherapy using vinorelbine (VNR) plus dihydropyrimidine dehydrogenase-inhibitory fluoropyrimidine (DIF) in EGFR-mutated lung adenocarcinomas, and we investigated a potential mechanism by which the combination chemotherapy of VNR + DIF was favorable in the treatment of EGFR-mutated lung adenocarcinoma in vitro. In our retrospective analysis, the response rate and disease control rate afforded by the VNR + DIF treatment tended to be better than those by platinum-based chemotherapy, and the progression-free survival of the 24 VNR + DIF-treated patients was significantly longer than that of the 15 platinum-based chemotherapy patients. In EGFR-mutated PC9 cells, VNR induced EGFR dephosphorylation at a clinically achievable concentration. 1BR3-LR cells, a line of fibroblast cells transfected with a mutant EGFR construct, were completely resistant to gefitinib in the medium containing 10% fetal bovine serum (FBS), whereas the sensitivity of these cells to gefitinib was increased in 0.5% FBS-containing medium. Similarly, the sensitivity of 1BR3-LR cells to VNR was increased when they were cultured in low-serum condition. In addition, sodium orthovanadate (Na3VO4) inhibited the EGFR dephosphorylation induced by VNR or gefitinib and suppressed the cell growth inhibition by these agents in PC9 cells. VNR and gefitinib showed synergistic cell growth inhibition in combination with 5-fluorouracil (5-FU) in PC9 cells. We propose that the EGFR dephosphorylation induced by VNR is related to cell growth inhibitory activity of VNR, and that this is one of the mechanisms of the synergistic effect of VNR + 5-FU in EGFR-mutated lung cancer cells. In conclusion, the combination chemotherapy of VNR + DIF may be a promising treatment for NSCLC patients with EGFR mutations.

Background The purpose of this study was to evaluate the accuracy of contrast-enhanced high resolution helical computed tomography (CT) for assessing locoregional staging of palpable Tl-2 invasive breast cancer. Methods: Helical CT studies of 156 lesions from 156 patients with invasive breast cancer before breast-conserving surgery were examined. A lesion was defined as positive if focal enhancement was detected by CT within 100 seconds after contrast material administration. After resection, tumors were histopathologically mapped and comparison made with the extent of contrast enhancement.

1988年から1998年4月までに当科で切除した胆嚢癌52例を臨床病理学的に検討し,そのリンパ節転移度から至適郭清範囲を検討した. m, mp症例は全例n0であり,胆嚢摘除術+D0-1で累積生存率100%と予後良好であった. ss以上32例におけるリンパ節転移陰性15例の累積生存率は73.1%で,陽性例の16.2%に比し有意に予後良好であった(p=0.0001).さらにss以上例をリンパ節転移度別に検討して見るとn0, n1症例の累積生存率は73.1%と予後良好でbinf0, hinf0であれば全層胆摘+D2で十分と考えられたがbinf, hinf陽性例の予後は不良で拡大手術が必要と考えられた. n2症例の累積生存率は20.8%と不良であったが,無再発生存例が2例あり,いずれもPpPD(PD)併施例で深達度ss, binf1以下, hinf2以下の症例であった. n3, n4症例は予後不良であった.現在当科ではn2(+)のss, se胆嚢癌においてhinf1-2, binf0-1であれば肝切除+PpPD(PD)+D3の良い適応と考え,胆嚢癌治療を行っている.

Background The optimum dose of granulocyte colony-stimulating factor (G-CSF) for peripheral blood stem cell (PBSC) mobilization after disease-oriented, conventional-dose chemotherapy remains unknown. Methods A multicenter dose-finding study of glycosylated G-CSF (lenograstim) for the mobilization of PBSCs following adjuvant CAF chemotherapy (cyclophosphamide, doxorubicin and 5-fluorouracil) was performed in 38 patients with postoperative breast cancer. Each 10, ten and eight patients were sequentially allocated to one of the three dose groups (2, 5 and 10 micrograms/kg, respectively) of lenograstim. Lenograstim was administered subcutaneously (s.c.) daily from day 8 to the day of the last apheresis and CD34+ cells and colony-forming units-granulocyte macrophage (CFU-GMs) in peripheral blood were measured serially. Additionally, 10 patients who received adjuvant CAF chemotherapy alone also participated in the study, as a control. Results Lenograstim was well tolerated up to 10 micrograms/kg, except for one patient given 10 micrograms/kg who developed transient grade 3 hepatic enzyme elevation. The peak levels of CD34+ cells and CFU-GMs in peripheral blood showed dose-response relationships. The median peak CD34+ cells for the 0, 2, 5 and 10 micrograms/kg dose groups were 5.4, 34.3, 55.0 and 127.6 cells/microliter, respectively, and those of CFU-GMs for the 0, 2, 5 and 10 micrograms/kg dose groups were 0.01, 0.33, 1.32 and 3.30 CFU-GMs/microliter, respectively. Conclusions Considering the previous reports suggesting that a pre-apheresis number of 40-50 CD34+ cells/microliter in peripheral blood is highly predictive for achievement of more than 2.5 x 10(6) CD34+ cells/kg in a standard apheresis procedure of 10 litres, the optimum dose of lenograstim for PBSC mobilization following CAF chemotherapy in patients with postoperative breast cancer is 5 micrograms/kg/day s.c.

Forty-seven cases of phyllodes tumor of the breast (resected and diagosed pathologicaly at this hospital from 1994 to August 1997) were evaluated clinicopathologically. Thirty-nine patients were classified as benign, 5 as borderline and 3 as malignant. Eighty-one percent of all tumors found were 5cm or less in diameter, and 38 tumors were 2cm or less in diameter. In this study, no clinical findings characteristic of benign or malignant tumors were identified. Some tumors needed to be differentiated from breast cancer. If an adequate surgical margin can be achieved, it would appear that local excision is an acceptable form of primary surgical treatment

Pancreatoduodenectomy (PD) has been applied for advanced gastric cancer with pancreatic and/or duodenal invasion. Twenty-one patients with gastric cancer underwent PD in our hospital from 1966 to 1996. No patients were lost to follow up. In this study, the clinicopathological factors and outcomes of these 21patients were analyzed. The numbers of patients with tumor invasion depths of T3 and T4 were 1 and 20, respectively. None had hepatic or peritoneal metastases, the exception being one patients who had adjacent peritoneal dissemination. Curative resection with extended lymphadenectomy beyond D2 was performed in all patients. The cumulative 1-year and 5-years survival rates were 65.2% and 41.5%, respectively. and the median survival time was 593days. There were five 5-year survivors. The survival rate in 5patients with positive lymph nodes on the posterior surface of the pancreatic head (No. 13 lymph node) was significantly lower than that in 16patients without No. 13 lymph node metastasis (p

Thirty seven patients with resectable peritoneal dissemination that were curability B in colorectal cancer were analyzed clinically and assessed factors which influence the survival. The 5-year survival rate was 31.4% and according to the multivariate analysis, there were two prognosis regulating factors included the number of disseminations of 3 and fewer and lymph node metastasis of n2 and under. Patients possess above two factors have good prognosis (survival at 5 years=56.8%). Based on prior statistical analysis. P1 and P2 divided into two groups according to the number of disseminations of 3 and fewer or 4 and over respectively. There were no statistical difference in two groups in P1. But in P2, three and fewer group had a statistically superior survival than 4 and over. And there were no statistical difference between 3 and fewer group of P2 and all of P1. These results suggest that it is appropriate that disseminations to the remole peritoneum of 3 and fewer are included P1 and 4 and over are included P2.

術前診断にて根治切除不可能と判定した進行胃癌21例に対し, 1993年より術前化学療法 (NAC) としてFLP療法 (CDDP 50mg/m2, day 1 day 8, 5FU 333mg/m2 and Leucovorin 30mg/body, day 1-8) を2クール以上施行してきた.19例に手術がなされ, 18例 (85.7%) が切除可能であった.【成績】(1) PR例は12例 (57.1%) 存在し, 根治Bとなった症例が7例存在した.(2) 部位別奏効率は, No.16リンパ節転移64.7%, 原発巣47.6%, 肝転移40%, 腹膜播種11.1%であった.(3) 21例の50%生存期間 (MST) は322日, 1生率40.5%であった.PR12例はMSTが471日で, 無効例は209日であった.(4) 根治度別では, 根治BのMST 835日に対し, 根治Cは310日で17.5か月MSTが延長していた.(5) 治療関連死亡, grade 4は1例もなく, grade 3以上の有害事象は白血球減少19.0%, 貧血14.3%, 口内炎9.5%などで安全性が確認された.【結語】FLP療法によるNACは, 非治癒因子がNo.16リンパ節転移である場合に治療効果が期待でき, PR例および根治B症例には生存率の向上も認められた.

Epidermal growth factor receptor (EGFR) is commonly overexpressed in malignant pleural mesothelioma (MPM). Cetuximab is a chimeric mouse-human antibody targeted against EGFR and induces potent antibody-dependent cellular cytotoxicity (ADCC). The action of cetuximab against MPM cells has not been well studied. Therefore, in this study, we investigated the antitumor activity of cetuximab against MPM cell lines, particularly with respect to ADCC activity in vitro and in vivo. EGFR expression of MPM cells was measured by a quantitative flow cytometric analysis and immunohistochemistry. The effect of cetuximab on growth inhibition was assessed using a modified MTT assay. The ADCC activity was measured by a 4-h 51Cr release assay using fresh or IL-2-activated peripheral blood mononuclear cells. In vivo antitumor activity of cetuximab was evaluated using an orthotopic implantation mouse model. Cetuximab-mediated ADCC activity against MPM cells was observed at low concentration (0.25 mg/ml) and was enhanced by IL-2, whereas no direct effect on growth inhibition was detected. A logarithmic correlation was observed between the number of EGFRs on MPM cells and ADCC activity. Low EGFR expression on the MPM cells, which was weakly detectable by immunohistochemistry, was sufficient for maximum ADCC activity. In the mouse model, cetuximab treatment with or without IL-2 significantly inhibited intrathoracic tumor growth and prolonged their survival. Our study shows that cetuximab has potent anti-MPM activity both in vitro and in vivo, mainly through the immunologic mechanism of ADCC. Cetuximab has the potential to be used as a novel therapy for MPM patients.

To elucidate whether adjuvant taxane monotherapy is a feasible and tolerable for postoperative breast cancer patients, we evaluated the severity of chemotherapy-induced peripheral neuropathy (CIPN) and the relative tolerability of regimens by health-related quality of life (HRQOL) assessment in node-positive breast cancer patients treated with taxane-containing regimens.We evaluated CIPN and HRQOL in the first 300 patients enrolled in a larger (1,060 total) multicenter phase III trial randomized to one of four adjuvant regimens: (1) anthracycline-cyclophosphamide followed by paclitaxel (ACP), (2) AC followed by docetaxel (ACD), (3) paclitaxel alone (PTX), or (4) docetaxel alone (DTX). CIPN was assessed by the Patient Neurotoxicity Questionnaire (PNQ) and the National Cancer Institute Common Toxicity Criteria, and HRQOL by Functional Assessment of Cancer Therapy-General (FACT-G). CIPN and HRQOL scores were compared between ACP and ACD vs. PTX and DTX, and ACP and PTX vs. ACD and DTX.PNQ sensory scores were significantly higher in patients treated with taxane monotherapy compared to treatment with AC followed by taxane (P = .003). No significant differences in PNQ sensory scores were observed between the ACP and PTX vs. ACD and DTX regimens (P = .669). Regardless of taxane regimen, PNQ severity scores for CIPN appear to be largely reversible within 1 year of adjuvant treatment. No significant difference in FACT-G scores was observed between any regimens during the study treatments.Patient-reported CIPN was significantly more severe with single-agent adjuvant taxane compared to AC followed by taxane treatment; however, the HRQOL findings support that single-agent taxane treatment is tolerable.

Elderly patients with advanced non-small cell lung cancer (NSCLC) require chemotherapy that is effective and minimally toxic. We evaluated the activity of a combination of vinorelbine and 5-fluorouracil (5-FU)/UFT (a fixed combination of tegafur and uracil) in vitro and in vivo to establish a rationale for clinical use. The cytotoxic activities of various combinations of vinorelbine and 5-FU, the active metabolite of tegafur, were analyzed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazlium bromide (MTT) assay and isobologram technique in vitro, using 3 NSCLC cell lines (A549, PC14, and Ma10). Sequential exposure to vinorelbine followed by 5-FU showed additive or synergistic activity against all 3 NSCLC cell lines tested. The reverse sequence showed no synergism. Antitumor activity and survival prolongation after treatment with different combinations of vinorelbine and UFT were evaluated in nude mice bearing PC14 xenografts. Treatment with vinorelbine before UFT was associated with higher antitumor activity, less toxicity, and longer survival than the reverse sequence. To clarify the underlying mechanism by which the combination exerts the synergistic effects, the expression of thymidylate synthase (TS) was assessed by Western blot analysis in vitro and by immunohistochemical analysis in an animal model. Vinorelbine suppressed the 5-FU-induced increase in TS protein in A549 cells. In PC14 tumor tissues of animal models, TS expression in cancer cells was suppressed by vinorelbine. Our data suggest that treatment with vinorelbine injection before oral UFT may have synergistic activity against NSCLC. This synergistic activity may be attributed to increased chemosensitivity to UFT caused by vinorelbine-induced suppression of TS.

Backgrounds: Bevacizumab (BEV) in combination with paclitaxel (PTX) and carboplatin (Cb) has provided survival benefit for advanced non-small cell lung cancer (NSCLC) as first-line treatment. However, tri-weekly PTX therapy induce severe peripheral neuropathy frequently, and may deteriorate quality of life. Because of this toxicity, platinum plus pemetrexed regimen may be preferred as first-line treatment over Cb/PTX/BEV regimen. Weekly paclitaxel (w-PTX) regimen has proved to be effective and safe in a large number of clinical trials for advanced solid tumors. However, few clinical studies have focused on the comparison between w-PTX and tri-weekly PTX in advanced NSCLC. We previously reported that fractionated administration of paclitaxel may be less toxic and more active against advanced NSCLC in clinical trials. Furthermore, several studies have reported that combination chemotherapy of w-PTX and BEV proved survival benefit in patients with advanced breast cancer. Therefore, we conducted a phase II trial to evaluate the efficacy and toxicity for the combination chemotherapy of w-PTX and BEV in patients with advanced non-squamous (non-Sq) NSCLC who have failed previous treatment. Methods: Eligibility required ECOG performance status 0-2 and prior cytotoxic chemotherapy. The primary end point of the phase II study was the progression free survival (PFS). PTX was infused on days 1, 8 and 15, and BEV (15 mg/kg) was administered on day 1 of a 4-week cycle until PD. Toxicity evaluations were based on CTCAE (version 4). Results: Between September 2011 and January 2014 a total of 30 eligible patients were enrolled onto the study. Conclusions: This combination of w-PTX and BEV is both feasible and active in the treatment of patients with advanced non-Sq NSCLC. We will soon start a phase II study evaluating this combination as first-line treatment.

Background: EGFR-tyrosine kinase inhibitors (TKI) often provides dramatic response to lung cancer harboring EGFR-activating mutations, but most patients eventually fail these treatment. We have reported the efficacy of vinorelbine (VNR) and dihydropyrimidine dehydrogenase (DPD) inhibitory fluoropyrimidine (DIF) for advanced non-small cell lung cancer patients, in particular EGFR mutant lung adenocarcinoma. The aim of this research was to elucidate the mechanism of efficacy of vinorelbine and DIF to EGFR mutant lung adenocarcinoma. Methods: We used a PC9 lung adenocarcinoma cell line with EGFR- activating mutation, and evaluated whether VNR or 5FU affect the phosphorylation of EGFR using Western blotting. Then, we transfected mutated EGFR gene to 1BR3 fibroblast cell line and measured the sensitivity of VNR to 1BR3 with or without transfection of mutated EGFR. Results: In PC9 cells, VNR induced dose- and time-dependent dephosphorylation of EGFR at clinically achievable concentrations, but 5-FU did not. EGF phosphorylated EGFR and reduced the sensitivity of PC9 cells to VNR. Sodium vanadate induced sustained phosphorylation of EGFR, and also conferred resistance to VNR. The sensitivity of VNR to 1BR3 cells transfected with mutated EGFR was higher than that of the parental 1BR3 cells. Conclusions: We showed that VNR inhibits proliferation of EGFR-mutated cells through EGFR dephosphorylation.

10624 Background: Hypoxia induces a wide spectrum of biological responses in tumors that alter tumor radio-sensitivity. In several tumors, including non-small cell lung carcinoma (NSCLC), the epidermal growth factor receptor (EGFR) is frequently over-expressed and activated in response to hypoxia, although the precise role of EGFR in hypoxia-associated radioresitance is unclear. Activating mutations in EGFR have been clinically linked to dramatic responses to EGFR tyrosine kinase inhibitors. We previously demonstrated that NSCLCs harboring activating EGFR mutations are radiosensitive with severe deficits in the non homologous end-joining (NHEJ) repair of DNA damage. Methods: We investigated hypoxia associated radiation responses in a panel of NSCLC cell lines and immortalized human bronchial epithelial cells expressing wild type EGFR (WT) or expressed EGFR forms with somatic activating L858R or DE746-E750 mutations (MT). Results: (1) Relative to WT EGFR, MT EGFR expression is associated with significantly higher radiosensitivity under chronic hypoxia (2) Under hypoxic conditions, MT EGFR expression is associated with a unique spectrum of DNA damage responses (DDR) that significantly deviated from canonical response of WT EGFR expressing NSCLCs (3) Genome wide gene expression analysis identified a dramatic hypoxia-associated DDR shift to selective down-regulation of homologous recombination (HR). Moreover, MT EGFR deficits in NHEJ caused a synthetic lethality effect in the context of hypoxia through a simultaneous blockade of HR and NHEJ. (4) Blockade of WT EGFR by the anti-EGFR monoclonal antibody, cetuximab perfectly recapitulated the contextual synthetic lethality of MT EGFR expression. (5) Cetuximab had a dramatic synergistic effect on survival of rat lung orthotopic tumors where hypo-fractionated radiotherapy or cetuximab alone had marginally controlled tumors. Conclusions: The data support a critical role for EGFR-mediated DNA repair in hypoxia associated tumor radio-resistance. Furthermore, this role could be a target for refractory hypoxic EGFR-WT tumor especially in the context of hypo-fractionated radiotherapy which, in its current form, is potentially counter-productive for hypoxic tumors.

6619 Background: Physician-based instruments (e.g., NCI-CTC) are widely used to assess chemotherapy-induced peripheral neuropathy (CIPN). However, current evidence suggests that physician-based assessments under-report the incidence and severity of CIPN. To overcome this limitation, a patient-based questionnaire, patient neurotoxicity questionnaire (PNQ) was developed, and a phase III randomized adjuvant trial of breast cancer (N-SAS BC 02; AC followed by PAC/DOC vs. PAC/DOC alone) has demonstrated that PNQ is reliable and sensitive and responsive instrument to assess CIPN (Shimozuma et al., SABCS 2004; #6037). We prospectively evaluated the reliability and sensitivity of PNQ in advanced or metastatic breast cancer treated by weekly administration of paclitaxel. Moreover, a questionnaire survey was conducted on physician perspectives regarding the assessment of CPIN in Japan. Methods: CIPN and QOL were prospectively assessed in thirty-five patients with advanced or metastatic breast cancer who received weekly paclitaxel (80–100 mg/m2/w). PNQ and FACT-Ntx subscale were compared to NCI-CTC. Assessments were conducted at baseline, 8 wk, 16 wk after starting treatments. A questionnaire was sent to physician who participated in N-SAS BC 02 to clarify their perspectives regarding the CPIN. Results: Average response rate of the instruments was 89%. Sensory PNQ scores correlated with sensory FACT-Ntx scores (r=0.51), and NCI-CTC scores (r=0.58). NCI-CTC scores mainly distributed between 0 and 1, while PNQ scores widely distributed. Follow-up study revealed that sensory CIPN assessed by PNQ appeared to be sensitive as compared to NCI-CTC. In clinician survey, 47 out of 61 physicians (77%) responded, and majority of them considered neurosensory symptoms as diagnostic hallmark for CIPN. However, for the justification for treatment delay, dose modification, or treatment cessation, most laid weight on functional impairment in patients with CIPN. Most (80%) rated PNQ is helpful in management of patients at risk for CIPN. Conclusions: This study confirmed that physicians tended to underestimate CIPN, and PNQ was a more reliable and valid instrument to assess CIPN with high acceptability in physicians. [Table: see text]

8523 Background: Chemotherapy-induced peripheral neuropathy (CIPN) commonly occurs during taxane chemotherapy (Tx). There is no standardized approach used in the assessment of CIPN. Physician-based instruments (e.g., NCI-CTC) are widely used for this purpose, but are associated with several important limitations. As current medical evidence suggests that physician-based assessments under-report the incidence and severity of subjective symptoms, they cannot reliably or accurately assess symptoms of CIPN. We prospectively assessed CIPN during Tx in a phase III randomized trial to evaluate the reliability and sensitivity of these different approaches. Methods: Three hundred breast cancer (BC) patients entered the National Surgical Adjuvant Study of Breast Cancer 02 (N-SAS BC 02), a phase III randomized trial comparing 4 cycles of AC followed by 4 cycles of Tx (paclitaxel [PAC] or docetaxel [DOC]) vs. 8 cycles of Tx alone in N+ BC patients after surgery. CIPN and QOL were secondary endpoints. Patient-based PNQ and FACT-Ntx were compared to the clinician-based NCI-CTC. Assessments were conducted at baseline, 3rd, 5th, 7th cycles, 8 months, and one year after starting adjuvant treatments. A linear mixed effect model was used to compare data in the two treatment groups across repeated cycles of treatment. Results: Average response rate of the instruments was > 90%. Sensory PNQ scores strongly correlated with sensory FACT-Ntx scores (r=0.70), while they weakly correlated (r=0.43) with NCI-CTC scores. NCI-CTC scores distributed between 0 and 2, while PNQ scores distributed between 0 and 4. Comparison of CIPN during the first 3 cycles showed that significantly higher overall incidence of sensory disturbance was observed in Tx compared with AC as assessed by both PNQ (PAC, P [Table: see text]

Rho GTPases play an essential role in the control of various cellular functions. Accumulating evidence suggests that RhoA overexpression contributes to human cancer development. However, the activation states of RhoA are poorly defined in cancer cells. In this study, we examined both the expression levels and the activation states of RhoA in various lung cancer cells by quantitative real-time reverse transcriptase-polymerase chain reaction and in vivo Rho guanine nucleotide exchange assay, respectively. Moreover, we dissected the signaling pathway from the cell surface receptors to RhoA using a broad-spectrum G protein coupled receptor (GPCR) antagonist, [D-Arg1,D-Trp5,7,9,Leu11]Substance P (SP), and a recently reported Galphaq/11-selective inhibitor, YM-254890. We found that RhoA was expressed highly in large cell carcinoma cells but only weakly in adenocarcinoma cells. The activation states of RhoA are considerably different from its expression profiles. We found that four of six small cell lung carcinoma (SCLC) cell lines exhibited a moderate to high activation rate of RhoA. The addition of [D-Arg1,D-Trp5,7,9,Leu11]SP reduced RhoA activity by almost 60% in H69 SCLC cells. The addition of YM-254890 had no effect on RhoA activity in H69 cells. Our results suggest that RhoA is activated in various lung cancer cells independent of its expression levels, and the high activation state of RhoA in SCLC cells mainly depends on a neuroendocrine peptide autocrine system which signals through Galpha12 coupled GPCR to RhoA. This study provides new insights into RhoA signaling in lung cancer cells and may help in developing novel therapeutic strategies against lung cancer.