Proteome analysis of human pancreatic ductal adenocarcinoma tissue using two-dimensional gel electrophoresis and tandem mass spectrometry for identification of disease-related proteins.
A comparative proteomic approach has been used to identify and analyze proteins related to pancreatic cancer. Proteomes of eight pairs of clinical pancreatic ductal adenocarcinoma (PDAC) tissue samples and samples of normal adjacent tissue were obtained by two-dimensional gel electrophoresis (2DE). Comprehensive analysis of proteins was focused on total protein spots for which there were statistical differences between the two groups. Proteins were identified by peptide mass fingerprinting with tandem mass spectrometry (MS-MS). Western blotting and immunohistochemistry (IHC) were also performed to verify the expression of some candidate proteins. Thirty protein spots were identified, including proteases, antioxidant proteins, signal-transduction proteins, calcium-binding proteins, structural proteins, chaperones, and others. Western blotting and IHC confirmed up-regulated expression of two candidate proteins, nucleotide diphosphatase kinase (NDPK) and annexin II, in tumorous tissues. These results suggest that combination of 2DE with MS is an effective strategy for discovery of differently expressed proteins in PDAC which may be molecular markers for diagnosis or therapeutic targets.
Tian
Rui
R
Department of Biliary-Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China.
Wei
Li-Ming
LM
Qin
Ren-Yi
RY
Li
Yan
Y
Du
Zhi-Yong
ZY
Xia
Wei
W
Shi
Cheng-Jian
CJ
Jin
Hong
H
D017306:Annexin A2 / Q000378:metabolism; D014408:Biomarkers, Tumor; D015153:Blotting, Western; D021441:Carcinoma, Pancreatic Ductal / Q000737:chemistry* / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000737:chemistry* / Q000473:pathology; D020543:Proteome / Q000378:metabolism*; D012720:Severity of Illness Index; D053719:Tandem Mass Spectrometry / Q000379:methods*
D017306:Annexin A2; D014408:Biomarkers, Tumor; D020543:Proteome; D009701:Nucleoside-Diphosphate Kinase