No conflicts of interest were declared.
Proteome of formalin-fixed paraffin-embedded pancreatic ductal adenocarcinoma and lymph node metastases†
Article first published online: 17 JAN 2012
Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
The Journal of Pathology
Volume 226, Issue 5, pages 756–763, April 2012
How to Cite
Naidoo, K., Jones, R., Dmitrovic, B., Wijesuriya, N., Kocher, H., Hart, I. R. and Crnogorac-Jurcevic, T. (2012), Proteome of formalin-fixed paraffin-embedded pancreatic ductal adenocarcinoma and lymph node metastases. J. Pathol., 226: 756–763. doi: 10.1002/path.3959
- Issue published online: 6 MAR 2012
- Article first published online: 17 JAN 2012
- Accepted manuscript online: 14 NOV 2011 04:13AM EST
- Manuscript Accepted: 4 NOV 2011
- Manuscript Revised: 23 SEP 2011
- Manuscript Received: 2 AUG 2011
Vol. 227, Issue 4, e8, Article first published online: 10 JUL 2012
- lymphatic metastasis;
- laser capture microdissection;
- FFPE proteomics
Pancreatic ductal adenocarcinoma (PDAC) is a major cause of cancer-related death, largely due to metastatic disease. To better understand PDAC metastatic spread and identify novel therapeutic targets, we analysed the proteome of primary tumours and matched lymph node (LN) metastases. As frozen specimens of metastatic lesions are scarce, we examined formalin-fixed paraffin-embedded (FFPE) tissues. This poses technical challenges because of the cross-linkages induced by fixation. Using laser capture microdissection (PALM system), we isolated malignant epithelia from seven FFPE primary PDAC tumours and matched LN metastases. Following dissection, samples were analysed in duplicate using Multidimensional Protein Identification Technology (MudPIT); this resulted in the identification of 1504 proteins, 854 of which were common to all samples analysed. Comparison of the obtained proteins with data from previous proteomics studies on pancreatic tissue, pancreatic juice, serum, and urine resulted in a less than 30% overlap, indicating that our study has substantially expanded the current database of proteins expressed in this malignancy. Statistical analysis further showed that 115/854 proteins (13.5%) were significantly differentially expressed (g-value ≥ 3.8). Two proteins, S100P and 14-3-3 sigma, with highly significant g-values were confirmed to be significantly differentially expressed (S100P: p = 0.05 and 14-3-3 sigma: p < 0.001) in a larger series of 55 cases of matched primary PDAC and LN metastases using immunohistochemistry. Thus, laser capture microdissection of FFPE tissue coupled with downstream proteomic analysis is a valid approach for the investigation of metastatic PDAC. This is the first study to establish and compare the protein composition of primary PDAC and matched LN metastases, and has resulted in the identification of several potential epithelial-specific therapeutic targets, including 14-3-3 sigma and S100P. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.