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Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study

BACKGROUND: Large cell neuroendocrine carcinoma (LCNEC) of the lung, a subtype of large cell carcinoma (LCC), is characterized by neuroendocrine differentiation that small cell lung carcinoma (SCLC) shares. Pre-therapeutic histological distinction between LCNEC and SCLC has so far been problematic,...

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Autores principales: Nomura, Masaharu, Fukuda, Tetsuya, Fujii, Kiyonaga, Kawamura, Takeshi, Tojo, Hiromasa, Kihara, Makoto, Bando, Yasuhiko, Gazdar, Adi F, Tsuboi, Masahiro, Oshiro, Hisashi, Nagao, Toshitaka, Ohira, Tatsuo, Ikeda, Norihiko, Gotoh, Noriko, Kato, Harubumi, Marko-Varga, Gyorgy, Nishimura, Toshihide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3178477/
https://www.ncbi.nlm.nih.gov/pubmed/21888658
http://dx.doi.org/10.1186/2043-9113-1-23
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author Nomura, Masaharu
Fukuda, Tetsuya
Fujii, Kiyonaga
Kawamura, Takeshi
Tojo, Hiromasa
Kihara, Makoto
Bando, Yasuhiko
Gazdar, Adi F
Tsuboi, Masahiro
Oshiro, Hisashi
Nagao, Toshitaka
Ohira, Tatsuo
Ikeda, Norihiko
Gotoh, Noriko
Kato, Harubumi
Marko-Varga, Gyorgy
Nishimura, Toshihide
author_facet Nomura, Masaharu
Fukuda, Tetsuya
Fujii, Kiyonaga
Kawamura, Takeshi
Tojo, Hiromasa
Kihara, Makoto
Bando, Yasuhiko
Gazdar, Adi F
Tsuboi, Masahiro
Oshiro, Hisashi
Nagao, Toshitaka
Ohira, Tatsuo
Ikeda, Norihiko
Gotoh, Noriko
Kato, Harubumi
Marko-Varga, Gyorgy
Nishimura, Toshihide
author_sort Nomura, Masaharu
collection PubMed
description BACKGROUND: Large cell neuroendocrine carcinoma (LCNEC) of the lung, a subtype of large cell carcinoma (LCC), is characterized by neuroendocrine differentiation that small cell lung carcinoma (SCLC) shares. Pre-therapeutic histological distinction between LCNEC and SCLC has so far been problematic, leading to adverse clinical outcome. We started a project establishing protein targets characteristic of LCNEC with a proteomic method using formalin fixed paraffin-embedded (FFPE) tissues, which will help make diagnosis convincing. METHODS: Cancer cells were collected by laser microdissection from cancer foci in FFPE tissues of LCNEC (n = 4), SCLC (n = 5), and LCC (n = 5) with definite histological diagnosis. Proteins were extracted from the harvested sections, trypsin-digested, and subjected to HPLC/mass spectrometry. Proteins identified by database search were semi-quantified by spectral counting and statistically sorted by pair-wise G-statistics. The results were immunohistochemically verified using a total of 10 cases for each group to confirm proteomic results. RESULTS: A total of 1981 proteins identified from the three cancer groups were subjected to pair-wise G-test under p < 0.05 and specificity of a protein's expression to LCNEC was checked using a 3D plot with the coordinates comprising G-statistic values for every two group comparisons. We identified four protein candidates preferentially expressed in LCNEC compared with SCLC with convincingly low p-values: aldehyde dehydrogenase 1 family member A1 (AL1A1) (p = 6.1 × 10(-4)), aldo-keto reductase family 1 members C1 (AK1C1) (p = 9.6x10(-10)) and C3 (AK1C3) (p = 3.9x10(-10)) and CD44 antigen (p = 0.021). These p-values were confirmed by non-parametric exact inference tests. Interestingly, all these candidates would belong to cancer stem cell markers. Immunohistochmistry supported proteomic results. CONCLUSIONS: These results suggest that candidate biomarkers of LCNEC were related to cancer stem cells and this proteomic approach via FFPE samples was effective to detect them.
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spelling pubmed-31784772011-09-23 Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study Nomura, Masaharu Fukuda, Tetsuya Fujii, Kiyonaga Kawamura, Takeshi Tojo, Hiromasa Kihara, Makoto Bando, Yasuhiko Gazdar, Adi F Tsuboi, Masahiro Oshiro, Hisashi Nagao, Toshitaka Ohira, Tatsuo Ikeda, Norihiko Gotoh, Noriko Kato, Harubumi Marko-Varga, Gyorgy Nishimura, Toshihide J Clin Bioinforma Research BACKGROUND: Large cell neuroendocrine carcinoma (LCNEC) of the lung, a subtype of large cell carcinoma (LCC), is characterized by neuroendocrine differentiation that small cell lung carcinoma (SCLC) shares. Pre-therapeutic histological distinction between LCNEC and SCLC has so far been problematic, leading to adverse clinical outcome. We started a project establishing protein targets characteristic of LCNEC with a proteomic method using formalin fixed paraffin-embedded (FFPE) tissues, which will help make diagnosis convincing. METHODS: Cancer cells were collected by laser microdissection from cancer foci in FFPE tissues of LCNEC (n = 4), SCLC (n = 5), and LCC (n = 5) with definite histological diagnosis. Proteins were extracted from the harvested sections, trypsin-digested, and subjected to HPLC/mass spectrometry. Proteins identified by database search were semi-quantified by spectral counting and statistically sorted by pair-wise G-statistics. The results were immunohistochemically verified using a total of 10 cases for each group to confirm proteomic results. RESULTS: A total of 1981 proteins identified from the three cancer groups were subjected to pair-wise G-test under p < 0.05 and specificity of a protein's expression to LCNEC was checked using a 3D plot with the coordinates comprising G-statistic values for every two group comparisons. We identified four protein candidates preferentially expressed in LCNEC compared with SCLC with convincingly low p-values: aldehyde dehydrogenase 1 family member A1 (AL1A1) (p = 6.1 × 10(-4)), aldo-keto reductase family 1 members C1 (AK1C1) (p = 9.6x10(-10)) and C3 (AK1C3) (p = 3.9x10(-10)) and CD44 antigen (p = 0.021). These p-values were confirmed by non-parametric exact inference tests. Interestingly, all these candidates would belong to cancer stem cell markers. Immunohistochmistry supported proteomic results. CONCLUSIONS: These results suggest that candidate biomarkers of LCNEC were related to cancer stem cells and this proteomic approach via FFPE samples was effective to detect them. BioMed Central 2011-09-03 /pmc/articles/PMC3178477/ /pubmed/21888658 http://dx.doi.org/10.1186/2043-9113-1-23 Text en Copyright ©2011 Nomura et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Nomura, Masaharu
Fukuda, Tetsuya
Fujii, Kiyonaga
Kawamura, Takeshi
Tojo, Hiromasa
Kihara, Makoto
Bando, Yasuhiko
Gazdar, Adi F
Tsuboi, Masahiro
Oshiro, Hisashi
Nagao, Toshitaka
Ohira, Tatsuo
Ikeda, Norihiko
Gotoh, Noriko
Kato, Harubumi
Marko-Varga, Gyorgy
Nishimura, Toshihide
Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study
title Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study
title_full Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study
title_fullStr Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study
title_full_unstemmed Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study
title_short Preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. An FFPE proteomic study
title_sort preferential expression of potential markers for cancer stem cells in large cell neuroendocrine carcinoma of the lung. an ffpe proteomic study
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3178477/
https://www.ncbi.nlm.nih.gov/pubmed/21888658
http://dx.doi.org/10.1186/2043-9113-1-23
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