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. 1999 Jul;45(1):105–111. doi: 10.1136/gut.45.1.105

Induction of TFF1 gene expression in pancreas overexpressing transforming growth factor α

M Ebert 1, J Hoffmann 1, C Haeckel 1, K Rutkowski 1, R Schmid 1, M Wagner 1, G Adler 1, H Schulz 1, A Roessner 1, W Hoffmann 1, P Malfertheiner 1
PMCID: PMC1727567  PMID: 10369712

Abstract

BACKGROUND/AIMS—Chronic pancreatitis is an inflammatory disease of the exocrine pancreas associated with extensive fibrosis, enlarged pancreatic ducts, acinar cell degeneration, and the formation of tubular complexes. The molecular and biochemical alterations associated with these histological changes are not kown. Generally, the new family of TFF peptides (formerly known as P-domain peptides or trefoil factors) is aberrantly expressed during chronic inflammatory diseases of the gastrointestinal tract.
METHODS—Using human pancreatic tissues obtained from patients with chronic pancreatitis and murine pancreatic tissues obtained from transgenic mice overexpressing transforming growth factor α (TGF-α), the expression and cellular distribution of TFF1 was analysed using northern blot analysis, polymerase chain reaction (PCR), and immunohistochemistry.
RESULTS—In the normal human pancreas, TFF1 was scarce, with only a few ducts exhibiting cytoplasmic TFF1 immunoreactivity. In contrast, human chronic pancreatitis tissue specimens exhibited strong TFF1 immunoreactivity in ductal cells, areas of ductal hyperplasia, and tubular complexes. Semiquantitative PCR analysis of TFF1 mRNA levels showed enhanced expression of TFF1 in the pancreas of patients with chronic pancreatitis. Furthermore, TFF1 mRNA levels were detectable in the pancreas in four of five transgenic mice overexpressing TGF-α. In contrast, four of five wild type mice did not exhibit a TFF1 mRNA transcript. In addition, while no specific TFF1 immunoreactivity was present in the pancreas of the wild type mice, ductal epithelial cells and duct-like tubular complexes in the pancreas of the transgenic mice overexpressing TGF-α exhibited pronounced TFF1 immunoreactivity.
CONCLUSIONS—Ductal cells and tubular complexes in pancreatic fibrosis express TFF1. As the 5'-flanking region of TFF1 contains an epidermal growth factor responsive enhancer region and the expression of epidermal growth factor and TGF-α is enhanced in pancreatic fibrosis, the enhanced expression of TFF1 in pancreatic fibrosis may be mediated by TGF-α.


Keywords: fibrosis; pS2; pancreatitis; expression; transforming growth factor α; TFF peptides

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Figure 1  .

Figure 1  

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TFF1 immunoreactivity in tubular complexes in pancreatic fibrosis. (A) Transgenic mice overexpressing transforming growth factor α. Strong TFF1 immunoreactivity was observed in the duct-like cells of tubular complexes (arrowheads). (B) Human chronic pancreatitis. TFF1 immunoreactivity was also present at the surface of these duct-like cells forming a central lumen (arrowheads). Original magnifications: A, 120 ×; B, 400 ×.

Figure 2  .

Figure 2  

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Reverse transcription polymerase chain reaction (PCR) analysis of TFF1 expression in the pancreas of wild type and transgenic mice. With cDNAs obtained from total RNA which had been extracted from the pancreas of wild type mice (wt; n = 4) and mice overexpressing transforming growth factor α (tg; n = 5), a band corresponding to TFF1 was observed in the pancreas in four of five transgenic mice. In contrast, only one wild type mouse exhibited the TFF1 transcript in the pancreas. TFF2 was expressed in all transgenic and wild type mice, whereas TFF3 expression was detectable in three of five transgenic mice. Parallel PCR amplification of the cDNAs using two primers specific for glyceraldehyde-3-phosphate dehydrogenase (GAP) confirmed the integrity of the cDNAs used. M, DNA marker.    

Figure 3  .

Figure 3  

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TFF1 immunoreactivity in chronic pancreatitis. Strong TFF1 immunoreactivity was present in the vast majority of pancreatic ducts (A) and areas of ductal hyperplasia also exhibited intense TFF1 immunoreactivity (B,C). However, not all ducts exhibited TFF1 immunoreactivity (B, arrowheads), and acinar and islet cells, as well as fibroblasts and inflammatory cells, were generally devoid of TFF1 immunoreactivity. Original magnifications: A, B, 120 ×; C, 400 ×.

Figure 4  .

Figure 4  

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(A) Reverse transcription polymerase chain reaction (PCR) analysis of TFF1 expression in the normal pancreas and in chronic pancreatitis. With cDNA obtained from total RNA extracted from normal pancreatic tissue samples (n = 3) and samples from patients with chronic pancreatitis (n =3), a band corresponding to TFF1 was present in all samples, as detected by polyacrylamide gel electrophoresis. Parallel PCR amplification of the cDNAs using two primers specific for β-actin confirmed the integrity of the cDNAs used. M, DNA marker. (B) Semiquantitative PCR analysis of TFF1 expression in chronic pancreatitis. Total RNA was extracted as previously described and PCR analysis was performed. A 15 µl portion of the amplification mixture was removed every five cycles from cycles 25 to 35. The samples were dot-blotted to Nylon membranes and probed using a [32P]dCTP random prime labelled TFF1 cDNA probe.

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