Induction of tissue transglutaminase by dexamethasone: its correlation to receptor number and transglutaminase-mediated cell death in a series of malignant hamster fibrosarcomas.
Treatment of the hamster fibrosarcoma cell lines (Met B, D and E) and BHK-21 hamster fibroblast cells with the glucocorticoid dexamethasone led to a powerful dose-dependent mRNA-synthesis-dependent increase in transglutaminase activity, which can be correlated with dexamethasone-responsive receptor numbers in each cell line. Increasing the number of dexamethasone-responsive receptors by transfection of cells with the HG1 glucocorticoid receptor protein caused an increase in transglutaminase activity that was proportional to the level of transfected receptor. In all experiments the levels of the tissue transglutaminase-mediated detergent-insoluble bodies was found to be comparable with increases in transglutaminase activity. Despite an increase in detergent-insoluble body formation, an increase in apoptosis as measured by DNA fragmentation was not found. Incubation of cells with the non-toxic competitive transglutaminase substrate fluorescein cadaverine led to the incorporation of this fluorescent amine into cellular proteins when cells were damaged after exposure to trypsin during cell passage. These cross-linked proteins containing fluorescein cadaverine were shown to be present in the detergent-insoluble bodies, indicating that the origin of these bodies is via activation of tissue transglutaminase after cell damage by trypsinization rather than apoptosis per se, since Met B cells expressing the bcl-2 cDNA were not protected from detergent-insoluble body formation. We describe a novel mechanism of cell death related to tissue transglutaminase expression and cell damage.
Johnson
T S
TS
Department of Life Sciences, Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS, U.K.
Scholfield
C I
CI
Parry
J
J
Griffin
M
M
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D006224:Cricetinae; D003907:Dexamethasone / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D005354:Fibrosarcoma / Q000201:enzymology* / Q000473:pathology*; D009374:Neoplasms, Experimental / Q000201:enzymology* / Q000473:pathology*; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D011503:Transglutaminases / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000893:Anti-Inflammatory Agents; D011965:Receptors, Glucocorticoid; C028697:dexamethasone receptor; D003907:Dexamethasone; D011503:Transglutaminases