Proteomic Analyses of Pancreatic Cyst Fluids

作者：Ke Eileen; Patel Bhavinkumar B; Liu Tiffany; Li Xin Ming; Haluszka Oleh; Hoffman John P; Ehya Hormoz; Young Nancy A; Watson James C; Weinberg David S; Nguyen Minhhuyen T; Cohen Steven J; Meropol Neal J; Litwin Samuel; Tokar Jeffrey L; Yeung Anthony T^*

来源：Pancreas, 2009, 38(2): E33-E42.

DOI：10.1097/mpa.0b013e318193a08f

• pancreatic cyst fluid
• biomarkers
• proteomic
• mucin
• S100
• CEACAM

Objectives: There are currently no diagnostic indicators that are consistently reliable, obtainable, and conclusive for diagnosing and risk-stratifying pancreatic cysts. Proteomic analyses were performed to explore pancreatic cyst fluids to yield effective diagnostic biomarkers.
Methods: We have prospectively recruited 20 research participants and prepared their pancreatic cyst fluids specifically for proteomic analyses. Proteomic approaches applied were as follows: (1) matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry peptidomics with LC/MS/MS (HPLC-tandem mass spectrometry) protein identification; (2) 2-dimensional gel electrophoresis; (3) GeLC/MS/MS (tryptic digestion of proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by LC/MS/MS).
Results: Sequencing of more than 350 free peptides showed that exopeptidase activities rendered peptidomics of cyst fluids unreliable; protein nicking by proteases in the cyst fluids produced hundreds of protein spots from the major proteins, making 2-dimensional gel proteomics unmanageable; GeLC/MS/MS revealed a panel of potential biomarker proteins that correlated with carcinoembryonic antigen (CEA).
Conclusions: Two homologs of amylase, solubilized molecules of 4 mucins, 4 solubilized CEA-related cell adhesion molecules (CEA-CAMs), and 4 S100 homologs may be candidate biomarkers to facilitate future pancreatic cyst diagnosis and risk-stratification. This approach required less than 40 mu L of cyst fluid per sample, offering the possibility to analyze cysts smaller than 1 cm in diameter.

• 出版日期2009-3