Changes in the expression of hepatocyte protein gene-products associated with adaptation of cells to primary culture.
Expression of hepatocyte protein gene-products changes as cells adapt to the environment of tissue culture. We examined such changes, using "giant" two-dimensional (2-D) gel electrophoresis. Over 80 cellular and secreted proteins, about 3% of the 2500 we examined, were found to change spontaneously during the 20-h culture period. The magnitude of these changes was estimated to range from 1.5- to nearly 100-fold. If dexamethasone is included in the culture medium, the overall production of secreted proteins is maintained, but in addition it exerts major influences on the expression of individual protein gene-products. Ten of the spontaneous changes are enhanced and eight are unaffected, but most are substantially retarded (32) or even reversed (27). There are also large inductions or repressions in 12 proteins that do not change spontaneously in culture. We have tentatively identified tyrosine aminotransferase, phosphoenolpyruvate carboxykinase, gamma-fibrinogen, and alpha 1-acid glycoprotein to be among those induced by dexamethasone. Evidently, many more changes occur as these cells adapt to tissue culture than was previously appreciated. These results emphasize the power of 2-D gel analysis in monitoring the protein phenotype of cells. We have also shown that glucocorticoids are important in maintaining the overall synthesis of secreted proteins and the protein phenotype of isolated hepatocytes in primary culture.
D000818:Animals; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000096:biosynthesis; D005786:Gene Expression Regulation; D007525:Isoelectric Focusing; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009961:Orosomucoid / Q000096:biosynthesis; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000096:biosynthesis; D014176:Protein Biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D014444:Tyrosine Transaminase / Q000096:biosynthesis
D009961:Orosomucoid; D003907:Dexamethasone; D005340:Fibrinogen; D014444:Tyrosine Transaminase; D010729:Phosphoenolpyruvate Carboxykinase (GTP)