CAXII Is a Sero-Diagnostic Marker for Lung Cancer

To develop sero-diagnostic markers for lung cancer, we generated monoclonal antibodies using pulmonary adenocarcinoma (AD)-derived A549 cells as antigens by employing the random immunization method. Hybridoma supernatants were immunohistochemically screened for antibodies with AMeX-fixed and paraffin-embedded A549 cell preparations. Positive clones were monocloned twice through limiting dilutions. From the obtained monoclonal antibodies, we selected an antibody designated as KU-Lu-5 which showed intense membrane staining of A549 cells. Based on immunoprecipitation and MADLI TOF/TOF-MS analysis, this antibody was recognized as carbonic anhydrase XII (CAXII). To evaluate the utility of this antibody as a sero-diagnostic marker for lung cancer, we performed dot blot analysis with a training set consisting of sera from 70 lung cancer patients and 30 healthy controls. The CAXII expression levels were significantly higher in lung cancer patients than in healthy controls in the training set (P<0.0001), and the area under the curve of ROC was 0.794, with 70.0% specificity and 82.9% sensitivity. In lung cancers, expression levels of CAXII were significantly higher in patients with squamous cell carcinoma (SCC) than with AD (P = 0.035). Furthermore, CAXII was significantly higher in well- and moderately differentiated SCCs than in poorly differentiated ones (P = 0.027). To further confirm the utility of serum CAXII levels as a sero-diagnostic marker, an additional set consisting of sera from 26 lung cancer patients and 30 healthy controls was also investigated by dot blot analysis as a validation study. Serum CAXII levels were also significantly higher in lung cancer patients than in healthy controls in the validation set (P = 0.030). Thus, the serum CAXII levels should be applicable markers discriminating lung cancer patients from healthy controls. To our knowledge, this is the first report providing evidence that CAXII may be a novel sero-diagnostic marker for lung cancer

Long-term disease-free survivor of metastatic large-cell neuroendocrine carcinoma of the lung treated with amrubicin and irinotecan

Shinichiro Ryuge1, Shi-Xu Jiang2, Mayuko Wada1, Ken Katono1, Maiko Iwasaki1, Akira Takakura1, Sakiko Otani1, Yuka Kimura1, Tomoya Fukui1, Masanori Yokoba1, Masaru Kubota1, Masato Katagiri1, Kazusige Hayakawa3, Noriyuki Masuda11Department of Respiratory Medicine, 2Department of Pathology, 3Department of Radiology, Kitasato University School of Medicine, Sagamihara, Kanagawa, JapanAbstract: Large-cell neuroendocrine carcinoma (LCNEC) is a relatively uncommon variant of non-small cell lung cancer. Since the biological characteristics of LCNEC are similar to those of small cell lung cancer, LCNEC is usually treated with chemotherapy regimens used for small cell lung cancer. However, the outcomes are usually dismal. Here, we report a patient with LCNEC (a metastasis to the brain). After whole brain irradiation, he received a combination of amrubicin and irinotecan chemotherapy, and has been relapse-free for two years. This treatment regimen may be beneficial for patients with advanced LCNEC. Keywords: large-cell neuroendocrine carcinoma, chemotherapy, amrubicin, irinotecan, lung cance

Prognostic significance of nestin expression in patients with resected non-small cell lung cancer treated with platinum-based adjuvant chemotherapy; relationship between nestin expression and epithelial to mesenchymal transition related markers.

Although adjuvant platinum-based chemotherapy (AC) has been shown to improve survival of patients with completely resected stage II and stage IIIA non-small cell lung cancer (NSCLC), its effect is limited. Nestin is a class VI intermediate filament protein expressed in neural stem cells and several cancer cells including NSCLC. In the present study, we aimed to determine its prognostic significance concerning survival in NSCLC patients receiving AC.Nestin expression in cancer cells was immunohistochemically studied in 90 patients with completely resected stage II and stage IIIA NSCLC treated with AC and its association with clinicopathologic parameters, including ABCG2, E-cadherin, and vimentin expression, was evaluated. Kaplan-Meier survival analysis and Cox proportional hazards models were used to estimate the effect of nestin expression on survival.Nestin expression was observed in 28 of the 90 (31.1%) NSCLCs. Clinicopathologically, nestin expression was associated with loss of E-cadherin expression (P = 0.006) and vimentin positive expression (P < 0.001). In survival analysis, nestin expression was significantly associated with a poorer prognosis (P = 0.028). Multivariable analysis confirmed that nestin expression is an independent prognostic indicator in NSCLC patients receiving AC (HR = 2.56; 95% CI, 1.23-5.30, P = 0.01).The present study reveals that nestin expression is a prognostic indicator of a poorer survival probability in NSCLC patients receiving AC, although its prognostic significance still requires confirmation with larger patient populations

Antibody absorption test for MYH9.

<p>KU-LU-6 antibody supernatant was pre-absorbed with none (A), 0.12 ug (B), 0.24 ug (C), and 0.48 ug (D) of synthetic MYH9 proteins. Each absorbed antibody was immunostained with formalin-fixed LC2/ad-cis cells. The stainability of KU-Lu-6 antibody was gradually reduced depending on the concentration of MYH9 protein.</p

Immunohistochemical screening of the Ku-Lu-6 antibody with AMeX-fixed LCN1-cis (A) and LCN1 (B) cells.

<p>Membranous staining was observed in LCN1-cis cells, but not in LCN1 cells. (original magnification: A, B ×400).</p

Characteristics of the Patients.

<p>AD = adenocarcinoma; NS = never smoker; p-TNM = pathological TNM; S = smoker</p><p>SQ = squamous cell carcinoma</p><p>*Each case was reassigned to a pathological stage on the basis of the IASLC Lung Cancer Staging Project (seventh edition).</p><p>Characteristics of the Patients.</p

Immunoprecipitation with KU-Lu-6 antibody.

<p>Lane 1: molecular weight marker, lane 2: LC-2/ad-cis lysate combined with KU-Lu-6 antibody, lane 3: LC-2/ad-cis lysate combined with protein L, lane 4: KU-Lu-6 antibody combined with protein L, lane 5: LC-2/ad-cis lysate. Lanes 3 and 4 are negative controls, and specific immunoprecipitated product with KU-Lu-6 antibody was detected in lane 2 (arrow).</p