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J Biol Chem, Vol. 273, Issue 13, 7495-7500, March 27, 1998

Up-regulation of Transforming Growth Factor (TGF)- Receptors by TGF-1 in COLO-357 Cells

From the Departments of Medicine, Biological Chemistry, and Pharmacology, University of California, Irvine, California 92697

In the present study we investigated the actions of transforming growth factor (TGF)-1 on gene induction and cyclin-dependent kinase inhibitors in relation to TGF- receptor modulation in COLO-357 pancreatic cancer cells. TGF-1 inhibited the growth of COLO-357 cells in a time- and dose-dependent manner and caused a rapid but transient increase in plasminogen activator inhibitor-I and insulin-like growth factor binding protein-3 mRNA levels. TGF-1 caused a delayed but sustained increase in the protein levels of the cyclin-dependent kinase inhibitors p15^Ink4B, p21^Cip1, and p27^Kip1 and a sustained increase in type I and II TGF- receptors (TRI and TRII) mRNA and protein levels. The protein synthesis inhibitor cycloheximide (10 µg/ml) completely blocked the TGF-1-mediated increase in TRI and TRII expression. Furthermore, a nuclear runoff transcription assay revealed that the increase in receptor mRNA levels was due to newly transcribed RNA. There was a significant increase in TRI and TRII mRNA levels in confluent cells in comparison to subconfluent (80% confluent) controls, as well as in serum- starved cells when compared with cells incubated in medium containing 10% fetal bovine serum. COLO-357 cells expressed a normal SMAD4 gene as determined by Northern blot analysis and sequencing. These results indicate that TGF-1 modulates a variety of functions in COLO-357 cells and up-regulates TGF- receptor expression via a transcriptional mechanism, which has the potential to maximize TGF-1-dependent antiproliferative responses.

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