Quantitative proteomic analysis of mitochondrial proteins differentially expressed between small cell lung cancer cells and normal human bronchial epithelial cells.

Abstract

BACKGROUND:

Small cell lung cancer (SCLC) is highly aggressive and is associated with a dismal prognosis. However, there are no clinically recognized biomarkers for early diagnosis. In this study, we used quantitative proteomics to build differential mitochondrial protein profiles that may be used for early diagnosis and investigated the pathogenesis of lung cancer.

METHODS:

We cultured SCLC cells (NCI-H446) and normal human bronchial epithelial cells (16-HBE); mitochondria were extracted and purified using differential and Percoll density gradient centrifugation. Subsequently, we used Western blot analysis to validate mitochondrial purity and labeled proteins/peptides from NCI-H446 and 16-HBE cells using relative and absolute quantification of ectopic tags. We then analyzed mixed samples and identified proteins using two-dimensional liquid chromatography-tandem mass spectrometry. Additionally, we performed subsequent bioinformatic proteome analyses using the programs ExPASy, GOA, and STRING. Finally, the relationship between ornithine aminotransferase expression and clinicopathological features in lung cancer patients was evaluated using immunohistochemistry.

RESULTS:

One hundred and fifty-three mitochondrial proteins were differentially expressed between 16-HBE and NCI-H446 cells. The expression of 30 proteins between 16-HBE and NCI-H446 cells increased more than 1.3-fold. The upregulation of ornithine aminotransferase was associated with pathological grade and clinical tumor node metastasis stage.

CONCLUSION:

Our experiment represented a promising method for building differential mitochondrial protein profiles between NCI-H446 and 16-HBE cells. Such analysis may also help to identify novel biomarkers of lung cancer.

• Authors: Wei Li, Wei Zhang, Wenjing Deng, Yujie Zhong, Yonghong Zhang, Zhuo Peng, Haijuan Chen, Ruiying Sun, Xuemei Zhang, Shuanying Yang
• Journal: Thoracic cancer (Thorac Cancer) Vol. 9 Issue 11 Pg. 1366-1375 (11 2018) ISSN: 1759-7714 [Electronic] Singapore
• PMID: 30198174 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd.

Chemical References

• Mitochondrial Proteins

Topics

• Bronchi (metabolism)
• Cell Line, Tumor
• Cell Proliferation
• Epithelial Cells (metabolism)
• Female
• Humans
• Lung Neoplasms (genetics, pathology)
• Male
• Mitochondrial Proteins (metabolism)
• Proteomics (methods)
• Small Cell Lung Carcinoma (genetics, pathology)