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S‐adenosylmethionine decarboxylase gene expression is regulated by the cAMP signal transduction pathway in H‐ras transformed fibrosarcoma cells capable of malignant progression
Author(s) -
Hurta Robert A. R.
Publication year - 2001
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.1108
Subject(s) - cycloheximide , forskolin , biology , cell culture , signal transduction , microbiology and biotechnology , gene expression , fibroblast , transfection , protein biosynthesis , gene , biochemistry , genetics
Abstract The hypothesis that H‐ras transformed cells contain alterations in signalling pathways important in controlling the expression of S‐adenosylmethionine decarboxylase, (SAMDC) a highly regulated activity in the biosynthesis of polyamines was tested. Mouse 10 T1/2 fibroblasts and H‐ras transformed cell lines of varying degrees of malignant potential were treated with agents which affect cAMP levels within cells. Elevations in SAMDC expression were noted in H‐ras transformed metastatic C3 cells, which were not observed in either parental, non‐transformed 10 T1/2 fibroblast cells, or in ras transformed NR3 cells, which are only capable of benign tumour formation. Forskolin, a stimulator of cAMP synthesis, was able to increase SAMDC enzyme activity but the response which occurred was dependent upon the cellular phenotype expressed. Actinomycin D pre‐treatment of C3 cells prior to exposure to forskolin did not abrogate the elevation observed in SAMDC gene expression suggesting that this was not a transcriptional process mediated event. Forskolin pre‐treatment of C3 cells did result in a marked increase in the half‐life of SAMDC mRNA transcripts suggesting a role for post‐transcriptional stabilization. Furthermore, cycloheximide treatment of malignant C3 cells resulted in elevated SAMDC mRNA levels. Treatment of malignant C3 cells with both cycloheximide and forskolin together resulted in a further additive elevation in SAMDC message levels. Cycloheximide treatment alone was found to affect the half‐life of SAMDC mRNA through a mechanism of post‐transcriptional stabilization. Additionally, altered SAMDC gene expression in C3 cells which occurred in response to cAMP alterations, was enhanced by stimulation of a protein kinase C pathway suggesting possible interactions between protein kinase C‐and cAMP‐mediated pathways which affect the regulation of SAMDC expression in highly malignant C3 cells. These results demonstrate aberrant regulation of signalling pathways involved in controlling SAMDC gene expression in H‐ras transformed cells capable of malignant progression and provide further insight into the altered growth regulatory program associated with H‐ras mediated cellular transformation and malignant progression. J. Cell. Biochem. Suppl. 36: 209–221, 2001. © 2001 Wiley‐Liss, Inc.