High prevalence of gene abnormalities in young patients with lung cancer.

BACKGROUND: Recently, driver oncogenes in adenocarcinoma of the lung were identified, and several molecular target agents were introduced in the clinical setting. However, there are few reports on the frequency of gene abnormalities in young patients with lung cancer. MATERIALS AND METHODS: Twelve patients with lung adenocarcinoma aged 40 or younger at Juntendo University Urayasu Hospital or Juntendo University Hospital from July 2004 to March 2010 were analyzed for driver oncogene status including EGFR activating mutation, EML4-ALK fusion gene, and K-ras mutation. RESULTS: Four patients showed EGFR gene mutation. Five out of 7 EGFR mutation-negative patients showed positive results for EML4-ALK gene fusion. One case whose EGFR mutation was indeterminate. CONCLUSIONS: Driver oncogene including EGFR mutation and EML4-ALK fusion gene was identified in 9 of 12 cases (75%). Examination of gene abnormalities is essential in young patients with non-small cell lung cancer to provide the best treatment.

Predictive factors associated with the success of pneumatic dilatation in Japanese patients with primary achalasia: a study using high-resolution manometry.

BACKGROUND/AIMS: A new classification of achalasia using high-resolution manometry (HRM) has recently been suggested. Pneumatic dilatation (PD) is a common treatment for primary achalasia. The usefulness of the new classification and HRM for the treatment and follow-up of patients after PD is unknown. The aim of this study was to evaluate the PD effectiveness and the predictive factors of success in Japanese patients with achalasia using HRM and the new classification of achalasia. METHODS: Twenty-five patients were diagnosed with primary achalasia using HRM and treated by PD in our hospital. We evaluated symptom scores and esophageal manometry 6 and 12 months after the first PD. RESULTS: After the first PD treatment, remission occurred in 24 out of 25 (96.0%) patients at 6 months and in 19 out of 25 (76.0%) patients at 12 months. With the new classification of achalasia, the success rates were 83.3, 80.0 and 50% for types I, II and III, respectively, 12 months after PD. The median age of the successful group was significantly greater than that of the failure group (47.1 vs. 37.0 years, p < 0.05). The median residual lower esophageal sphincter (LES) pressure 6 months after PD in the successful group was significantly lower than that of the failure group (9.0 vs. 15.5 mm Hg, p < 0.05). CONCLUSION: Good predictors of PD success were old age (>40 years) and residual LES pressures less than 15 mm Hg 6 months after PD.

Hydration with magnesium and mannitol without furosemide prevents the nephrotoxicity induced by cisplatin and pemetrexed in patients with advanced non-small cell lung cancer.

BACKGROUND: The aim of this study was to examine the effect of hydration with magnesium and mannitol without furosemide on the nephrotoxocity accompanying combination chemotherapy using cisplatin and pemetrexed in patients with advanced non-small cell lung cancer (NSCLC). METHODS: Fifty patients with NSCLC who received cisplatin plus pemetrexed, using either old hydration protocol including normal saline with mannitol and furosemide, or a new one including normal saline with magnesium and mannitol without furosemide were retrospectively analyzed. Nephrotoxicity was compared between patients treated using the old protocol and those treated with the new protocol. Univariate and multivariate analyses were performed to identify the independent factors associated with protection against nephrotoxicity in patients with NSCLC who received cisplatin plus pemetrexed. RESULTS: Thirty patients received the old hydration protocol, while 20 patients were treated using the new hydration protocol. The patients treated using the new hydration protocol showed a significantly greater increase in creatinine clearance (P=0.0004) and a decrease in the serum creatinine level (P=0.0148) after one course of chemotherapy compared with those treated using the old hydration protocol. There were no differences in the chemotherapeutic response or overall survival between the groups (P=0.572). The new hydration protocol with supplemented magnesium with mannitol without furosemide was an independent factor for the protection against nephrotoxicity induced by cisplatin and pemetrexed in patients with advanced NSCLC [HR 0.232 (95% CI: 0.055-0.986), P=0.039]. CONCLUSIONS: These results demonstrate that the new hydration protocol comprising supplementation with magnesium without furosemide could prevent the nephrotoxicity induced by cisplatin and pemetrexed without affecting the treatment outcome.

[Efficacy of erlotinib after gefitinib administration in patients with non-small cell lung cancer].

We retrospectively evaluated the survival benefit of dispensing erlotinib after gefitinib administration in patients with nonsmall cell lung cancer. Ninety patients treated with erlotinib in our hospital were divided into two groups: G+ group patients who were treated with erlotinib with prior gefitinib administration, and G- group patients who were treated with erlotinib without prior gefitinib administration. Median survival time (MST) in all 90 patients was 275 days. MST of 22 patients in the G+ group was shorter than that of 68 patients in G- group, but this difference was not statistically significant (283 days vs 177 days, p=0. 329). MST in 19 patients of the G+group who were administered erlotinib for over 1 month was shorter than that of 49G-group patients who were administered erlotinib over 1 month. However, this difference was also not statistically significant(395 days vs 238 days, p=0. 575). Univariate analysis demonstrated that EGFR mutation unknown, time to progression (TTP) with gefitinib longer than 1 year, gefitnib administration longer than 1 year, and responder to gefitinib, suggest a better prognosis. Mutivariate analysis revealed that only TTP with gefitinib longer than 1 year was an independent prognostic factor for patients in the G+ group.

Combined pH-impedance monitoring and high-resolution manometry of Japanese patients treated with proton-pump inhibitors for persistent symptoms of non-erosive reflux disease.

BACKGROUND: Data on acid and non-acid reflux patterns and esophageal function in Japanese patients with non-erosive reflux disease (NERD) are limited. The aim of this study was to use combined multichannel intraluminal impedance pH monitoring (MII-pH) and high-resolution manometry (HRM) to investigate the characteristics of Japanese patients who were treated with a "double-dose" (20 mg) of rabeprazol (a proton-pump inhibitor; PPI) for persistent symptoms of NERD. METHODS: Twenty-five patients who complained of typical gastroesophageal reflux disease symptoms, which had occurred more than twice a week despite treatment with rabeprazol, were enrolled in the study. All patients underwent upper endoscopy, esophageal HRM, and 24-h MII-pH monitoring while double-dose PPI therapy was continued. RESULTS: Twelve (48.0%) of the patients had a positive symptom index (SI) with 234 recorded symptoms, 127 (54.3%) of which were related to reflux episodes. Of those with reflux episodes, 29 (22.8%) were related to acid reflux, while 98 (77.2%) were the result of a weaker acidic reflux. In acid reflux and in mixed (liquid-gas) reflux, the proximal esophageal region was involved to a significantly greater degree (P<0.002 and P=0.005, respectively) than the distal region. In liquid reflux, there was no difference between the distal and proximal regions. HRM showed that proximal motility parameters were significantly more defective than in those of healthy volunteers. CONCLUSIONS: MII-pH monitoring indicated that weakly acidic reflux and mixed refluxate in the proximal esophagus is the major cause of persistent symptoms in patients with NERD who are being treated with PPI. HRM showed that proximal esophageal dysfunction might be a key condition that facilitates reflux.

Breakthrough lung Scedosporium prolificans infection with multiple cavity lesions in a patient receiving voriconazole for probable invasive aspergillosis associated with monoclonal gammopathy of undetermined significance (MGUS).

Breakthrough non- Aspergillus mold infections among patients receiving the anti-mold azole antifungal agents like voriconazole or posaconazole have been increasingly reported. We report a case of lung Scedosporium prolificans infection with multiple cavities in a 58-year-old man with monoclonal gammopathy of undetermined significance (MGUS) during voriconazole treatment for probable invasive aspergillosis. Cultures of repeated sputum specimens yielded the same fungus until his death 83 days after diagnosis. S. prolificans should be considered in patients with breakthrough infections receiving voriconazole.

[Twelve cases of advanced thymic carcinoma: a clinical review].

Thymic carcinoma is comparatively rare and no standard treatment has been established for advanced stage cases. We reviewed our therapeutic experience in 12 cases of thymic carcinoma. They consisted of 9 men and 3 women, ranging from 38 to 69 years of age, with a mean age of 56.5. According to Masaoka's classification, 5 cases were stage III and 7 were preoperative stage IVb. Nine cases were squamous cell carcinoma and 3 were adenocarcinoma. Four cases of preoperative clinical stage III underwent extended thymectomy, but none were completely resected and were classified as stage IV postoperatively. Chemotherapy combined with radiation therapy was given in 1 case, chemotherapy (monotherapy) was given in 4 cases, radiation therapy was given in 1 case, and 2 cases received best supportive care. The median survival time (MST) of patients who had undergone combined modality treatments including surgery was 1971 days, which was longer than the MST of 567 days of patients who were not able to undergo surgery. The prognosis outcome of advanced thymic carcinoma is poor, but combined modality therapies such as surgery, chemotherapy and radiation can be effective for some advanced thymic carcinoma cases.

Distinct roles of cholinergic receptors in small cell lung cancer cells.

BACKGROUND: Cholinergic receptors are expressed in small cell lung cancer (SCLC); however, the distinct functions of muscarinic cholinergic receptor 3 (mAChR3) and the nicotinic cholinergic receptor (nAChR) in SCLC have not yet been completely elucidated. MATERIALS AND METHODS: RT-PCR and Western blotting were used to investigate the expression of cholinergic receptors. Flow cytometry was used to detect the integrin expression. Cell proliferation, adhesion and migration assays were carried out in vitro to determine the roles of the cholinergic receptors in SBC3 human SCLC cells. RESULTS: Both mAChR3 and nAChR were expressed in the SBC3 cells. Acetylcholine iodide (Ach) stimulated SBC3 cell proliferation, adhesion and migration toward fibronectin (Fn). The mAChR3 antagonist, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), or the nAChR antagonist, mecamylamine hydrochloride (Meca), inhibited SBC3 cell proliferation in the presence or the absence of exogenous Ach. 4-DAMP abrogated cell adhesion and migration toward Fn induced by Ach, while Meca had no effect. Interestingly, Ach did not alter Fn receptor (alphavbeta1 or alpha5beta1 integrin) expression, while anti-beta1 integrin antibody or anti-alphav and anti-alpha5 integrin antibody completely abrogated cell adhesion to Fn induced by Ach. CONCLUSION: Both mAChR3 and nAChR are expressed in SCLC. SBC3 cell proliferation is regulated in vitro through both cholinergic receptors. In contrast, SBC3 cell migration and adhesion toward Fn are modulated only by mAChR. Moreover, the stimulatory effects of Ach on cell adhesion and migration through mAChR3 are presumably modulated by functional alteration of alphavbeta1 and alpha5beta1 integrin, but not by any variation in their expression. The mAChR3 antagonist may therefore be a beneficial therapeutic modality for SCLC patients, especially those with chronic obstructive pulmonary disease (COPD) as a comorbidity.

Osteopontin modulates malignant pleural mesothelioma cell functions in vitro.

BACKGROUND: Although serum osteopontin (OPN) concentration is elevated in patients with malignant pleural mesothelioma (MPM), the role of OPN in the pathogenesis and development of MPM remains unknown. MATERIALS AND METHODS: To determine the roles of OPN in MPM, immunohistochemical staining was performed to investigate the concentration of OPN in the pleural tumor of patients with mesothelioma; cell adhesion, proliferation and migration assays of H28 cells, an MPM cell line, were also carried out in vitro. RESULTS: H28 cells cultured on OPN-coated plates revealed enhanced adhesion, proliferation, migration, cell survival and phosphorylated focal adhesion kinase activities. As expected, these enhancements were markedly suppressed with the addition of anti-alphavbeta3 antibody or arginine-glycine-aspartic acid serine (RGDS) peptide to the medium. CONCLUSION: OPN is speculated to play an important role in the enhancement of adhesion, proliferation and migration activities of H28 cells, presumably by interacting with the alphavbeta3 integrin.

Osteopontin is involved in the development of acquired chemo-resistance of cisplatin in small cell lung cancer.

Osteopontin (OPN) is a multi-functional cytokine involved in cell survival, migration and adhesion which is associated with tumorigenesis, progression and metastasis. However, the role of OPN in chemo-sensitivity of human lung cancer has not yet been elucidated. The purpose of this study is to investigate the role of OPN in chemo-sensitivity of lung cancer cells. We developed a stable OPN transfectant (SBC-3/OPN) and a control transfectant (SBC-3/NEO) from human small cell lung cancer cell line, SBC-3. SBC-3/OPN cells were more resistant to cisplatin than SBC-3/NEO cells. Multi-drug resistance-associated protein (MRP) does not appear to be involved in the development of acquired chemo-resistance, since MRP inhibitor did not alter chemo-sensitivity. After exposure to cisplatin, the apoptotic SBC-3/OPN cells were reduced in number compared to SBC-3/NEO cells. Treatment with cisplatin revealed that the expression of anti-apoptotic protein, bcl-2, was down-regulated in SBC-3/NEO cells, while that of SBC-3/OPN cells was not altered. In contrast, pro-apoptotic protein, bax, was not altered in both SBC-3/OPN and SBC-3/NEO cells, thus bcl-2/bax ratio was decreased in SBC-3/NEO but not altered in SBC-3/OPN cells. Activation of caspase-3 and caspase-9 was increased in SBC-3/NEO cells, but not in SBC-3/OPN cells. Our results suggest that OPN enhances chemo-resistance of cisplatin in SBC-3 cells by suppressing bcl-2 protein down-regulation, thereby blocking the caspase-9- and caspase-3-dependent cell apoptosis.

Osteopontin is involved in the formation of malignant pleural effusion in lung cancer.

Malignant pleural effusion (MPE) is associated with advanced-stage lung cancer and is a poor prognostic sign for these patients. Osteopontin (OPN) is a multifunctional cytokine that is involved in the tumor progression and angiogenesis of lung cancer cells. The purpose of this study is to investigate and provide evidence for the role of OPN in the formation of MPE associated with lung cancer. In this study, we established an OPN knockdown murine lung cancer cell line, 3LL cells, utilizing the small interfering RNA (siRNA) technique. To reveal the effect of OPN on the formation of MPE associated with lung cancer, we directly injected OPN knockdown 3LL cells, 3LL/OPN siRNA, or control cells, 3LL/control siRNA, into the pleural space of C57BL/6 mice. OPN knockdown significantly reduced the formation of MPE, but did not inhibit in vivo tumor growth of 3LL cells in mice. Vascular endothelial growth factor (VEGF) concentration in MPE was markedly decreased in the 3LL/OPN siRNA in comparison with that of the 3LL/control siRNA. In vitro, recombinant OPN protein enhanced VEGF secretion from human umbilical vein endothelial cell (HUVEC) or human mesothelial cell line, Met5A cells, in a concentration-dependent manner. These results suggest that OPN is positively involved in the formation of MPE of lung cancer presumably by promoting VEGF secretion from vascular endothelial cells or mesothelial cells. OPN could be an effective target molecule for reducing MPE in lung cancer patients.

Abrogation of the interaction between osteopontin and alphavbeta3 integrin reduces tumor growth of human lung cancer cells in mice.

Osteopontin (OPN) is a multifunctional cytokine involved in cell signaling by interacting with alphavbeta3 integrins. Recent clinical studies have indicated that OPN expression is associated with tumor progression and poor prognosis among patients with lung cancer. However, the biological role of OPN in human lung cancer has not yet been well-defined. The purpose of this study is to investigate and provide evidence for the causal role of OPN regarding tumor growth and angiogenesis in human lung cancer. In this study, we developed a stable OPN transfectant from human lung cancer cell line SBC-3 which does not express the intrinsic OPN mRNA. To reveal the in vivo effect of OPN on tumor growth of human lung cancer, we subcutaneously injected OPN-overexpressing SBC-3 cells (SBC-3/OPN) and control cells (SBC-3/NEO) into the nude mice. Transfection with the OPN gene significantly increased in vivo tumor growth and neovascularization of SBC-3 cells in mice. These in vivo effects of OPN were markedly suppressed with administration of anti-alphavbeta3 integrin monoclonal antibody or anti-angiogenic agent, TNP-470. Furthermore, recombinant OPN protein enhanced human umbilical vein endothelial cell (HUVEC) proliferation in vitro, and this enhancement was significantly inhibited with the addition of anti-alphavbeta3 integrin antibody. Taken together, these results suggest that OPN plays a crucial role for tumor growth and angiogenesis of human lung cancer cells in vivo by interacting with alphavbeta3 integrin. Targeting the interaction between OPN and alphavbeta3 integrin could be effective for future development of anti-angiogenic therapeutic agents for patients with lung cancer.

Signal transduction mediated by endostatin directly modulates cellular function of lung cancer cells in vitro.

Endostatin (ED) is a carboxyl-terminal fragment of collagen XVIII with strong antiangiogenic activity. ED has been considered as a highly specific inhibitor of endothelial cell proliferation and migration through interaction with its receptor on the surface of endothelial cells. Recently, direct antitumor effects of ED in colon cancer cells and head and neck squamous cell carcinoma cells has been reported. However, its effect on lung cancer cells has not been clarified. The purpose of the present study was to determine the effect of ED on in vitro lung cancer cell function and to identify its receptor on lung cancer cells. We revealed that alpha5 integrin is capable of being a functional ED receptor among several integrins that are expressed on murine lung cancer (Lewis lung cancer [LLC]) cells. We further demonstrated that the ED-integrin interaction modulates various in vitro biological functions of LLC cells as we revealed that immobilized ED helps in LLC cell adhesion and migration in an integrin-dependent manner. Furthermore, ED inhibited LLC cell proliferation and induced apoptosis. Interestingly, ED did not demonstrate any antiproliferative activity against the other murine lung cancer cell line, KLN205, that lacks alpha5 integrin but binds to immobilized ED through the beta1 integrin. In addition, the binding of ED to alpha5 integrin on LLC cells induced phosphorylation of focal adhesion kinase. Taken together, these results suggest that the interaction between ED and alpha5 integrin may play an important role in lung cancer cell function.

Interaction between CD44 and hyaluronate induces chemoresistance in non-small cell lung cancer cell.

CD44s is a principle hyaluronate (HA) receptor and has been reported to play an important role in cancer cell invasion and metastasis. The aim of our study is to determine if the interaction between HA and CD44s influences in vitro chemosensitivity of non-small cell lung cancer (NSCLC). NSCLC cell line, H322 cells, transfected with the CD44s gene (H322/CD44s) cultured on HA coated plates were more resistant to cisplatin (CDDP) than that on bovine serum albumin. Multidrug resistance protein2 (MRP2) expression was induced in H322/CD44s cells cultured on HA. MRP2 inhibitor, MK571, not only suppressed MRP2 expression but also reversed CDDP resistance. These results suggest that the interaction between CD44s and HA play a pivotal role in acquired resistance to CDDP in NSCLC and MRP2 could be involved in this potential mechanism.

Prognostic factors in patients with inoperable non-small cell lung cancer--an analysis of long-term survival patients.

A survival benefit of patients with inoperable non-small cell lung cancer has been reported since a development of new therapeutic agents in the 1990's. However, multivariate analyses of patients have not been evaluated. The aim of this study is to identify prognostic factors in the long-term survivors who had been treated with chemotherapy using these new agents and/or radiotherapy. A retrospective study and clinical analysis of 121 inoperable nonsmall cell lung cancer patients were conducted. Fifteen cases (male: 9, female: 6) with a survival of more than 2 years were revealed. Regarding clinical variables between the 15 cases and others, an early nodal (N) status, a high serum protein level, a good performance status (PS) and those having first-line chemotherapy or radiotherapy were all identified as significant prognostic factors for the long-term survivors. Multivariate analyses also revealed that an early N status, a good PS, female gender and chemotherapy and/or radiotherapy were associated with the long-term survivors. These results suggest that patients with inoperable non-small cell lung cancer should be considered for appropriate treatments including new chemotherapeutic agents.

[Case of pulmonary mycobacterium avium complex disease, showing hypersensitivity pneumonitis-like diffuse shadow].

A 59-year-old man who had just completed therapy for tuberculosis, fell down in sauna and was admitted to a hospital. As acid-fast bacilli were positive (Gaffky 2) in sputum and residual cavity was shown in the right upper lobe on chest X-ray, he was transferred to our hospital. In spite of starting antituberculous chemotherapy, small nodular shadows appeared diffusely and were changed into ground-glass appearance on chest X-ray. The trans-bronchial-lung-biopsy revealed alveolitis mainly composed of lymphocyte infiltration with non-caseous epithelioid cell granulomas and organization which are likely to appear in hypersensitivity pneumonitis. As the acid-fast bacilli were identified as Mycobacterium avium, clarithromycin and kanamycin were added to the chemotherapy, but no improvement was observed in clinical feature. Corticosteroid therapy was further added and clinical feature improved immediately. Although we did not examine the presence of Mycobacterium avium in the water of sauna bath, we suspected this case as Hot Tub Lung based on clinical features and the response to treatment.

[Late onset hemiparkinsonism-hemiatrophy syndrome: a case report].

Hemiparkinsonism-hemiatrophy syndrome (HPHA) is a rare form of parkinsonism, characterized by unilateral parkinsonism, ipsilateral body atrophy and early age of onset. We report a 59-year-old woman with hemiatrophy of her face and upper limb on the left side presenting progressive hemiparkinsonism. Most of the HPHA patients have early age of onset, however, HPHA patients with age of onset over 50 years old have been reported. The clinical features of CBD are partly similar to those of HPHA, therefore it is important to consider HPHA as differential diagnosis of CBD even if the onset is late. In our patient, hemiatrophy was useful to differentiate HPHA from CBD. In previous reports, there was a difference between the dopamine D2 receptor binding capacity of CBD and that of HPHA. While the dopamine D2 receptor binding capacity of CBD was decreased asymmetrically, that of HPHA was not decreased. The PET finding of our patient revealed that asymmetrical reduction of [11C]-CFT uptake, meaning unilateral dopaminergic presynaptic hypofunction. However, [11C]-raclopride uptake, which assesses dopamine D2 receptor binding capacity, was normal. These PET findings are consistent with previous reports on HPHA. In our patient, hemiatrophy and PET findings were useful to diagnose HPHA.

Serum tenascin-C as a potential predictive marker of angiogenesis in non-small cell lung cancer.

BACKGROUND: Tenascin (Tn)-C is an extracellular matrix protein that is involved in tissue interactions during fetal development and oncogenesis. However, the role of serum Tn-C in non-small cell lung cancer (NSCLC) has not been clarified. PATIENTS AND METHODS: In this study, we determined the serum levels of Tn-C among NSCLC patients who underwent surgery, as well as other factors implicated for angiogenesis, to address the clinical implications in NSCLC. RESULTS AND CONCLUSION: The median concentration of serum Tn-C in NSCLC patients was slightly higher than that of normal controls, but this difference was not statistically significant. There was a positive correlation between serum Tn-C levels and microvessel density (MVD), serum osteopontin (OPN) and vascular endothelial growth factor (VEGF). In contrast, there was no correlation between serum Tn-C levels and serum carcinoembryonic antigen (CEA) and sialyl lewis-X (SLX) levels. The overall survival of patients with low Tn-C levels (<96 ng/ml) was significantly greater than that of patients with high Tn-C levels (> or =96 ng/ml). Intratumoral Tn-C expression was co-localized with expression of microvessels in the stroma of the cancer cells by immunohistochemical analysis. Moreover, enhanced in vitro migration of human umbilical vascular endothelial cells (HUVEC) was induced by recombinant Tn-C. Collectively, Tn-C may play an important role in angiogenesis of patients with NSCLC, and the determination of serum Tn-C may be useful in predicting intratumoral vasculature and patients' prognosis.