TRANSFORMING GROWTH FACTOR-BETA(1) SELECTIVELY REGULATES ORNITHINE DECARBOXYLASE GENE-EXPRESSION IN MALIGNANT H-RAS TRANSFORMED FIBROSARCOMA CELL-LINES

Negative growth regulators such as the transforming growth factor beta (TGF-beta) family appear to be important inhibitors in most tissue types. However, inhibition of DNA synthesis and cell proliferation is frequently lost during malignant transformation, and in some cases, tumor cell proliferation is actually stimulated by TGF-beta. The present study demonstrates a novel link between alterations in TGF-beta regulation during malignant conversion, and the expression of ornithine decarboxylase, a key rate-limiting activity in the biosynthesis of polyamines, and an enzyme that plays an important role in cell growth and differentiation. A panel of radiation and H-ras transformed mouse 10T 1/2 cell lines exhibiting increasing malignant potential was investigated for possible TGF-beta1 mediated changes in ornithine decarboxylase gene expression. Selective induction of gene expression was observed since only H-ras transformed cell lines with malignant potential exhibited marked elevations in ornithine decarboxylase message levels. Ornithine decarboxylase gene expression in nontransformed 10T 1/2 cells and cell lines capable of only benign tumor formation was unaffected by TGF-beta1 treatment. H-ras transformed cells were transfected with a plasmid placing the TGF-beta1 coding region under the control of a zinc sensitive metallothionein promoter. When these cells were cultured in the presence of zinc an elevation of TGF-beta1 mRNA was observed within 30 min. This increase in TGF-beta1 message closely coincided with an elevation in ornithine decarboxylase message, and preceded an induction of jun-B, an early response gene in cells sensitive to TGF-beta1 stimulation. Evidence for regulation of ornithine decarboxylase gene expression by TGF-beta1 at both transcription and posttranscription was found. Actinomycin D pretreatment of malignant cells prior to TGF-beta1 exposure prevented the increase in ornithine decarboxylase message. Marked differences in the rates of ornithine decarboxylase message decay were observed when cells treated with TGF-beta1 were compared to untreated controls, with the half-life of ornithine decarboxylase mRNA increasing from 2.5 h in untreated cells to 17.5 h in cells exposed to TGF-beta1. In addition, evidence was obtained for a cycloheximide sensitive regulator of ornithine decarboxylase gene expression, since the presence of this protein synthesis inhibitor increased the levels of ornithine decarboxylase message, and this effect was synergistically augmented by exposure of cells to cycloheximide and induction of TGF-beta1 gene expression together. These results show for the first time that TGF-beta1 can regulate ornithine decarboxylase expression in malignant H-ras transformed cells, and suggest a mechanism of growth factor stimulation of malignant cells, in which early alterations in the control of ornithine decarboxylase, gene expression are important. (C) 1993 Wiley-Liss, Inc.