Proteome analysis of gastric cancer metastasis by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of metastasis-related proteins.
A well-described animal model was used to understand the molecular mechanisms of carcinogenesis and metastasis of gastric cancer at the protein level. Gastric cancer was induced in 12 Wistar rats by oral administration of N-methyl-N'-Nitro-N-Nitrosoguanidine (MNNG). The protein expression patterns of normal gastric tissue, gastric cancer, and corresponding metastases were analyzed by proteomics in matched tissues of 3 rats. Proteins in the region of molecular masses of 15-75 kDa and an isoelectric point of 3-7 were separated by two-dimensional electrophoresis (2-DE) and identified by peptide fingerprinting with matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS). Twenty-seven spots corresponding to 25 different proteins served as landmarks for comparison between tissues. The identified proteins included cytoskeletal proteins, stress associated proteins, proteins involved in signal transduction, cell proliferation and differentiation, and metabolism. Eleven proteins were up-regulated and 2 proteins were down-regulated in tumor tissue when compared with normal tissue. Twelve proteins were up-regulated and 8 proteins were down-regulated in the metastases when compared with the primary tumor. The overexpression of HSP27 in gastric cancer was confirmed by immunohistochemical analysis of human gastric cancer specimens. Combining well-defined animal models with proteome analysis will improve our understanding of the fundamental changes that contribute to the process of carcinogenesis and the formation of metastases in gastric cancer.
Department of Gastroenterology, Hepatology and Infectious Diseases, Research Center Immunology/Institute of Experimental Internal Medicine, and Pathology, Otto-von-Guericke University, Magdeburg, Germany.
Jie
J
Chen
Thilo
T
Kähne
Christoph
C
Röcken
Tobias
T
Götze
Jun
J
Yu
Joseph J Y
JJ
Sung
Minhu
M
Chen
Pinjin
P
Hu
Peter
P
Malfertheiner
Matthias P A
MP
Ebert
D000230:Adenocarcinoma / Q000737:chemistry / Q000556:secondary; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D008114:Liver Neoplasms, Experimental / Q000737:chemistry / Q000556:secondary*; D008175:Lung Neoplasms / Q000737:chemistry / Q000556:secondary*; D008207:Lymphatic Metastasis; D008297:Male; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008875:Middle Aged; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000032:analysis; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013270:Stomach / Q000737:chemistry / Q000187:drug effects / Q000473:pathology; D013274:Stomach Neoplasms / Q000139:chemically induced / Q000737:chemistry / Q000473:pathology*; D014357:Trypsin / Q000378:metabolism; D015854:Up-Regulation
D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D011506:Proteins; D020543:Proteome; D008769:Methylnitronitrosoguanidine; D014357:Trypsin