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Published on: January 3, 2013
CD109 Overexpression in Pancreatic Cancer Identified by Cell-Surface Glycoprotein Capture
Randy S Haun1, Chun-Yang Fan2, Samuel G Mackintosh3
1Central Arkansas Veterans Healthcare System, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA ; Departments of Pharmaceutical Sciences, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA.
Background:
The development of novel targeted cancer therapies and/or diagnostic tools is dependent upon an understanding of the differential expression of molecular targets between normal tissues and tumors. Many of these potential targets are cell-surface receptors; however, our knowledge of the cell-surface proteins upregulated in pancreatic tumors is limited, thus impeding the development of targeted therapies for pancreatic cancer. To develop new diagnostic and therapeutic tools to specifically target pancreatic tumors, we sought to identify cell-surface proteins that may serve as potential tumor-specfic targets.
Methods:
Membrane glycoproteins on the pancreatic cancer cell lines BxPC-3 were labeled with the bifunctional linker biocytin hydrazide. Following proteolytic digestion, biotinylated glycopeptides were captured with streptavidin-coupled beads then released by PNGaseF-mediated endoglycosidase cleavage and identified by liquid chromatography-tandem mass spectrometry (MS). A protein identified by the cell-surface glycoprotein capture procedure, CD109, was evaluated by western analysis of lysates of pancreatic cancer cell lines and by immunohistochemistry in sections of pancreatic ductal adenocarcinoma and non- neoplastic pancreatic tissues.
Results:
MS/MS analysis of glycopeptides captured from BxPC-3 cells revealed 18 proteins predicted or known to be associated with the plasma membrane, including CD109, which has not been reported in pancreatic cancer. Western analysis of CD109 in lysates prepared from pancreatic cancer cell lines revealed it was expressed in 6 of 8 cell lines, with a high level of expression in BxPC-3, MIAPaCa-2, and Panc-1 cells. Immunohistochemical analyses of human pancreatic tissues indicate CD109 is significantly overexpressed in pancreatic tumors compared to normal pancreas.
Conclusions:
The selective capture of glycopeptides from the surface of pancreatic cancer cell lines can reveal novel cell-surface glycoproteins expressed in pancreatic ductal adenocarcinomas.
Insights
Researchers identified CD109 as a novel cell-surface protein overexpressed in pancreatic tumors. This discovery aids in developing targeted therapies and diagnostic tools for pancreatic cancer.
Area of Science:
- Oncology
- Proteomics
- Molecular Biology
Background:
- Targeted cancer therapies require understanding molecular differences between normal and tumor tissues.
- Limited knowledge of cell-surface proteins in pancreatic tumors hinders targeted therapy development.
- Identifying novel cell-surface targets is crucial for advancing pancreatic cancer diagnostics and therapeutics.
Purpose of the Study:
- To identify novel cell-surface proteins overexpressed in pancreatic tumors.
- To discover potential targets for new pancreatic cancer diagnostic and therapeutic tools.
Main Methods:
- Utilized biocytin hydrazide labeling of pancreatic cancer cell lines (BxPC-3).
- Employed streptavidin-coupled beads for capturing biotinylated glycopeptides.
- Identified proteins using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
- Validated CD109 expression via Western analysis and immunohistochemistry.
Main Results:
- Identified 18 plasma membrane-associated proteins, including CD109, a novel finding in pancreatic cancer.
- CD109 was expressed in 6 out of 8 pancreatic cancer cell lines, notably high in BxPC-3, MIAPaCa-2, and Panc-1.
- Immunohistochemistry confirmed significant CD109 overexpression in pancreatic tumors compared to normal pancreatic tissue.
Conclusions:
- Selective capture of cell-surface glycoproteins effectively identifies novel targets in pancreatic cancer.
- CD109 represents a promising novel target for pancreatic ductal adenocarcinoma therapies and diagnostics.
