Identification of major cellular proteins synthesized in response to interleukin-1 and interleukin-6 in human hepatoma HepG2 cells.
Interleukin-1 (IL-1) and interleukin-6 (IL-6) are principal proinflammatory cytokines inducing the acute phase response of various tissues, including liver. Cultured human hepatoma HepG2 cells were stimulated with IL-1 (10 ng/ml) and IL-6 (10 ng/ml). After 24 h the cells were collected and disrupted by sonication in a lysis buffer containing 8M urea. The extracted cellular proteins were separated by 2D polyacrylamide gel electrophoresis. The gels were stained with Coomassie Brilliant Blue R-250 and the protein spots showing different intensities in comparison to control (unstimulated) cells were excised and subjected to analysis by LC-MS/MS. Alternatively, proteins were stained with SYPRO Ruby. These differentially expressed proteins include seven up-regulated and two down-regulated intracellular proteins of various functions. The identification of three cytokine-responsive proteins was confirmed by biosynthetic labeling with [35S]methionine after incubation of HepG2 cells, and by western blot with specific antisera.
Bonar
Emilia
E
Faculty of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland.
Dubin
Adam
A
Bierczyńska-Krzysik
Anna
A
Noga
Marek
M
Silberring
Jerzy
J
Stalińska
Krystyna
K
Koj
Aleksander
A
D051339:Actin Depolymerizing Factors / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins / Q000378:metabolism; D044927:Electron-Transferring Flavoproteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D050556:Fatty Acid-Binding Proteins / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000494:pharmacology*; D008113:Liver Neoplasms / Q000473:pathology; D008565:Membrane Proteins / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D011506:Proteins / Q000378:metabolism*; D013460:Sulfur Isotopes; D013482:Superoxide Dismutase / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects*
D051339:Actin Depolymerizing Factors; D003598:Cytoskeletal Proteins; D044927:Electron-Transferring Flavoproteins; D050556:Fatty Acid-Binding Proteins; D007375:Interleukin-1; D053583:Interleukin-1beta; D015850:Interleukin-6; D008565:Membrane Proteins; D011506:Proteins; D013460:Sulfur Isotopes; C071776:radixin; D008715:Methionine; D013482:Superoxide Dismutase