CAXII Is a sero-diagnostic marker for lung cancer.

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 paraffi...

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Autores principales: Makoto Kobayashi, Toshihide Matsumoto, Shinichiro Ryuge, Kengo Yanagita, Ryo Nagashio, Yoshitaka Kawakami, Naoki Goshima, Shi-Xu Jiang, Makoto Saegusa, Akira Iyoda, Yukitoshi Satoh, Noriyuki Masuda, Yuichi Sato
Formato: article
Lenguaje:EN
Publicado: Public Library of Science (PLoS) 2012
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Medicine
R
Science
Q
Acceso en línea:https://doaj.org/article/f42a5fdb1ecd4432afc4c81d59c4e9e3
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spelling oai:doaj.org-article:f42a5fdb1ecd4432afc4c81d59c4e9e32021-11-18T07:24:52ZCAXII Is a sero-diagnostic marker for lung cancer.1932-620310.1371/journal.pone.0033952https://doaj.org/article/f42a5fdb1ecd4432afc4c81d59c4e9e32012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22439015/?tool=EBIhttps://doaj.org/toc/1932-6203To 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.Makoto KobayashiToshihide MatsumotoShinichiro RyugeKengo YanagitaRyo NagashioYoshitaka KawakamiNaoki GoshimaShi-Xu JiangMakoto SaegusaAkira IyodaYukitoshi SatohNoriyuki MasudaYuichi SatoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 3, p e33952 (2012)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Makoto Kobayashi
Toshihide Matsumoto
Shinichiro Ryuge
Kengo Yanagita
Ryo Nagashio
Yoshitaka Kawakami
Naoki Goshima
Shi-Xu Jiang
Makoto Saegusa
Akira Iyoda
Yukitoshi Satoh
Noriyuki Masuda
Yuichi Sato
CAXII Is a sero-diagnostic marker for lung cancer.
description 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.
format article
author Makoto Kobayashi
Toshihide Matsumoto
Shinichiro Ryuge
Kengo Yanagita
Ryo Nagashio
Yoshitaka Kawakami
Naoki Goshima
Shi-Xu Jiang
Makoto Saegusa
Akira Iyoda
Yukitoshi Satoh
Noriyuki Masuda
Yuichi Sato
author_facet Makoto Kobayashi
Toshihide Matsumoto
Shinichiro Ryuge
Kengo Yanagita
Ryo Nagashio
Yoshitaka Kawakami
Naoki Goshima
Shi-Xu Jiang
Makoto Saegusa
Akira Iyoda
Yukitoshi Satoh
Noriyuki Masuda
Yuichi Sato
author_sort Makoto Kobayashi
title CAXII Is a sero-diagnostic marker for lung cancer.
title_short CAXII Is a sero-diagnostic marker for lung cancer.
title_full CAXII Is a sero-diagnostic marker for lung cancer.
title_fullStr CAXII Is a sero-diagnostic marker for lung cancer.
title_full_unstemmed CAXII Is a sero-diagnostic marker for lung cancer.
title_sort caxii is a sero-diagnostic marker for lung cancer.
publisher Public Library of Science (PLoS)
publishDate 2012
url https://doaj.org/article/f42a5fdb1ecd4432afc4c81d59c4e9e3
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