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

Journal of Clinical Bioinformatics

Toshihide Nishimura⁷  Gyorgy Marko-Varga⁵  Harubumi Kato²  Noriko Gotoh⁸  Norihiko Ikeda^1,10  Tatsuo Ohira^1,10  Toshitaka Nagao¹  Hisashi Oshiro¹  Masahiro Tsuboi^1,10  Adi F Gazdar^1,11  Yasuhiko Bando⁶  Makoto Kihara⁷  Hiromasa Tojo³  Takeshi Kawamura⁴  Kiyonaga Fujii⁹  Tetsuya Fukuda⁶  Masaharu Nomura¹ 

[1] Diagnostic Pathology, Division, Tokyo Medical University, Tokyo, Japan;Niizashiki Central General Hospital, Saitama, Japan;Dept. of Biophysics and Biochemistry, Osaka University, Graduate School of Medicine, Suita, Japan;Laboratory for Systems Biology and Medicine, RCAST, The University of Tokyo, Tokyo, Japan;Clinical Protein Science & Imaging, Dept. of Measurement Technology and Industrial Electrical Engineering, Lund University, Lund, Sweden;Biosys Technologies, Inc., Tokyo, Japan;Medical ProteoScope Co., Ltd. Tokyo, Japan;Division of Systems Biomedical Technology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan;Dept. of Structural Biology, Graduate School of Pharmaceutical Science. Hokkaido, University, Hokkaido, Japan;Dept. of Surgery I, Tokyo Medical University, Tokyo, Japan;Hamon Center for Therapeutic Cancer Research, UT Southwestern Medical Center, Texas, USA

关键词: cancer stem cell markers;    mass spectrometry;    formalin-fixed paraffin embedded tissues;    large cell neuroendocrine carcinoma;   

Others  :  806566
DOI  :  10.1186/2043-9113-1-23

 received in 2011-04-01, accepted in 2011-09-03,  发布年份 2011

【 摘 要 】

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.

【 授权许可】

   
2011 Nomura et al; licensee BioMed Central Ltd.

【 图 表 】

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