Limits...
Extensive quantitative remodeling of the proteome between normal colon tissue and adenocarcinoma.

Wiśniewski JR, Ostasiewicz P, Duś K, Zielińska DF, Gnad F, Mann M - Mol. Syst. Biol. (2012)

Bottom Line: Functionally similar changes in the proteome were observed comparing rapidly growing and differentiated CaCo-2 cells.In contrast, there was minimal proteomic remodeling between primary cancer and metastases, suggesting that no drastic proteome changes are necessary for the tumor to propagate in a different tissue context.Our proteomic data set furthermore allows mapping quantitative changes of functional protein classes, enabling novel insights into the biology of colon cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Proteomics and Signal Transduction, Max-Planck-Institute of Biochemistry, Martinsried, Germany. jwisniew@biochem.mpg.de

ABSTRACT
We report a proteomic analysis of microdissected material from formalin-fixed and paraffin-embedded colorectal cancer, quantifying > 7500 proteins between patient matched normal mucosa, primary carcinoma, and nodal metastases. Expression levels of 1808 proteins changed significantly between normal and cancer tissues, a much larger fraction than that reported in transcript-based studies. Tumor cells exhibit extensive alterations in the cell-surface and nuclear proteomes. Functionally similar changes in the proteome were observed comparing rapidly growing and differentiated CaCo-2 cells. In contrast, there was minimal proteomic remodeling between primary cancer and metastases, suggesting that no drastic proteome changes are necessary for the tumor to propagate in a different tissue context. Additionally, we introduce a new way to determine protein copy numbers per cell without protein standards. Copy numbers estimated in enterocytes and cancer cells are in good agreement with CaCo-2 and HeLa cells and with the literature data. Our proteomic data set furthermore allows mapping quantitative changes of functional protein classes, enabling novel insights into the biology of colon cancer.

Show MeSH

Related in: MedlinePlus

Immunohistochemical staining of adenocarcinoma (A, D, G) and normal mucosa (B, E, H) with antibodies against PALM3, MFI2, and GPR56. (C, F, I) Statistical evaluation of the staining of 20 sample pairs. All differences between cancer and normal were significant at P<0.0001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3472694&req=5

f4: Immunohistochemical staining of adenocarcinoma (A, D, G) and normal mucosa (B, E, H) with antibodies against PALM3, MFI2, and GPR56. (C, F, I) Statistical evaluation of the staining of 20 sample pairs. All differences between cancer and normal were significant at P<0.0001.

Mentions: Proteins that are significantly upregulated in cancer samples can be considered as potential biomarkers. Among these, cell-surface proteins are particularly attractive because they may be targeted by antibodies. Table III lists 34 upregulated plasma membrane proteins. Two low abundant and one medium abundant protein were selected for further validation by immunohistochemistry. Antibodies against PALM3, MFI, and GPR56 strongly stained cancer cells whereas only weak staining was observed in normal tissue, thus validating upregulation of the proteins (Figure 4). Knowledge on proteins significantly changed between normal tissue and adenocarcinoma can be used for subsequent targeted analysis of larger number of samples, using either IHC or mass spectrometry-based methods. This may identify clinically relevant correlations between protein upregulation in cancer and the disease outcome. Another attractive follow-up to this study would be to search for the proteins identified as outliers as secreted proteins in plasma samples (Hanash et al, 2008). In this way, clinically useful biomarkers could be identified and potentially later monitored by conventional ELISA.


Extensive quantitative remodeling of the proteome between normal colon tissue and adenocarcinoma.

Wiśniewski JR, Ostasiewicz P, Duś K, Zielińska DF, Gnad F, Mann M - Mol. Syst. Biol. (2012)

Immunohistochemical staining of adenocarcinoma (A, D, G) and normal mucosa (B, E, H) with antibodies against PALM3, MFI2, and GPR56. (C, F, I) Statistical evaluation of the staining of 20 sample pairs. All differences between cancer and normal were significant at P<0.0001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3472694&req=5

f4: Immunohistochemical staining of adenocarcinoma (A, D, G) and normal mucosa (B, E, H) with antibodies against PALM3, MFI2, and GPR56. (C, F, I) Statistical evaluation of the staining of 20 sample pairs. All differences between cancer and normal were significant at P<0.0001.
Mentions: Proteins that are significantly upregulated in cancer samples can be considered as potential biomarkers. Among these, cell-surface proteins are particularly attractive because they may be targeted by antibodies. Table III lists 34 upregulated plasma membrane proteins. Two low abundant and one medium abundant protein were selected for further validation by immunohistochemistry. Antibodies against PALM3, MFI, and GPR56 strongly stained cancer cells whereas only weak staining was observed in normal tissue, thus validating upregulation of the proteins (Figure 4). Knowledge on proteins significantly changed between normal tissue and adenocarcinoma can be used for subsequent targeted analysis of larger number of samples, using either IHC or mass spectrometry-based methods. This may identify clinically relevant correlations between protein upregulation in cancer and the disease outcome. Another attractive follow-up to this study would be to search for the proteins identified as outliers as secreted proteins in plasma samples (Hanash et al, 2008). In this way, clinically useful biomarkers could be identified and potentially later monitored by conventional ELISA.

Bottom Line: Functionally similar changes in the proteome were observed comparing rapidly growing and differentiated CaCo-2 cells.In contrast, there was minimal proteomic remodeling between primary cancer and metastases, suggesting that no drastic proteome changes are necessary for the tumor to propagate in a different tissue context.Our proteomic data set furthermore allows mapping quantitative changes of functional protein classes, enabling novel insights into the biology of colon cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Proteomics and Signal Transduction, Max-Planck-Institute of Biochemistry, Martinsried, Germany. jwisniew@biochem.mpg.de

ABSTRACT
We report a proteomic analysis of microdissected material from formalin-fixed and paraffin-embedded colorectal cancer, quantifying > 7500 proteins between patient matched normal mucosa, primary carcinoma, and nodal metastases. Expression levels of 1808 proteins changed significantly between normal and cancer tissues, a much larger fraction than that reported in transcript-based studies. Tumor cells exhibit extensive alterations in the cell-surface and nuclear proteomes. Functionally similar changes in the proteome were observed comparing rapidly growing and differentiated CaCo-2 cells. In contrast, there was minimal proteomic remodeling between primary cancer and metastases, suggesting that no drastic proteome changes are necessary for the tumor to propagate in a different tissue context. Additionally, we introduce a new way to determine protein copy numbers per cell without protein standards. Copy numbers estimated in enterocytes and cancer cells are in good agreement with CaCo-2 and HeLa cells and with the literature data. Our proteomic data set furthermore allows mapping quantitative changes of functional protein classes, enabling novel insights into the biology of colon cancer.

Show MeSH
Related in: MedlinePlus