Transforming growth factor-beta 1 induces transforming growth factor-alpha promoter activity and transforming growth factor-alpha secretion in the human colon adenocarcinoma cell line FET

• Published in: Cancer research (Chicago, Ill.) Ill.), 1993-09, Vol.53 (17), p.4041-4047
• Main Authors: Lynch, M J, Pelosi, L, Carboni, J M, Merwin, J, Coleman, K, Wang, R C, Lin, P F, Henry, D L, Brattain, M G
• Format: Article
• Language: English
• Subjects: activity; adenocarcinoma; Adenocarcinoma - metabolism; Base Sequence; cell lines; colon; Colonic Neoplasms - metabolism; Culture Media, Serum-Free; Enzyme Induction; Humans; induction; Luciferases - biosynthesis; Luciferases - genetics; man; Molecular Sequence Data; Promoter Regions, Genetic; promoters; secretion; Transfection; Transforming Growth Factor alpha - biosynthesis; Transforming Growth Factor alpha - chemistry; Transforming Growth Factor alpha - genetics; Transforming Growth Factor beta - pharmacology; transforming growth factor- alpha; transforming growth factor- beta 1; Tumor Cells, Cultured
• ISSN: 0008-5472

FET cells are well differentiated human adenocarcinoma cells whose growth is partially inhibited (50-60%) by transforming growth factor-beta 1 (TGF-beta 1). In exponentially growing cultures, TGF-beta 1 induces the expression of transforming growth factor-alpha (TGF-alpha) by 3-fold. To determine whether this induction is the result of increased TGF-alpha promoter activity, FET cells were transiently transfected with a plasmid containing 2816 base pairs of the 5'-flanking region of the TGF-alpha gene linked to luciferase. Transfected FET cells treated with growth-inhibitory concentrations of TGF-beta 1 (10 ng/ml) showed up to a 10-fold increase in luciferase activity. The increase in luciferase activity was dose dependent through the normal physiological range of TGF-beta 1 (0.5-20 ng/ml), saturating at 10 ng/ml. This effect was also TGF-alpha promoter specific, inasmuch as the Rous sarcoma virus long terminal repeat used as a control remained relatively insensitive to the effects of TGF-beta 1. By using progressively smaller portions of the TGF-alpha promoter region, the TGF-beta 1-responsive element was mapped between base pairs -77 and -201 of the 5'-flanking region. TGF-beta 1 treatment also affected epidermal growth factor receptor levels. FET cells treated with TGF-beta 1 (10 ng/ml) for 48 h showed a 20% decrease in the number of epidermal growth factor receptors and a 2-fold increase in the number of high affinity epidermal growth factor receptors on their surface. These results indicate that TGF-beta 1 acts as a positive regulator of TGF-alpha transcription, and they suggest a possible mechanism by which these cells circumvent the growth-inhibitory effects of TGF-beta 1.