Pancreatitis-associated protein inhibits human pancreatic stellate cell MMP-1 and -2, TIMP-1 and -2 secretion and RECK expression.
OBJECTIVE
Pancreatic stellate cells (PSCs) play a key role in fibrogenesis associated with acute and chronic pancreatitis. Pancreatitis-associated protein (PAP), an acute-phase protein, is dramatically upregulated during acute and chronic pancreatitis. Assuming a protective role of PAP, we investigated its effects on human PSCs.


METHODS
PSCs were obtained by outgrowth from fibrotic human pancreas tissue. PAP was expressed in the yeast Pichia pastoris. PAP was added at 10 ng/ml to cultured PSCs. Cell proliferation was determined by bromodeoxyuridine incorporation. PSC migration was assessed by a wound healing assay. Collagen types I and III, fibronectin, matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs) and reversion-inducing cysteine-rich protein with Kazal motifs (RECK) were demonstrated on protein and mRNA level.


RESULTS
PAP had no significant effect on PSC proliferation and migration. Cell-associated fibrillar collagen types I and III and fibronectin increased after addition of PAP to PSCs. PAP diminished the expression of MMP-1 and -2 and TIMP-1 and -2 and their concentrations in PSC supernatants. RECK was detected on the surface of PSCs and its expression was reduced after PAP application.


CONCLUSIONS
Our data offer new insights into the biological functions of PAP, which may play an important role in wound healing response and cell-matrix interactions.
Li
Ling
L
Department of Clinical Chemistry, University Hospital Ulm, Ulm, Germany.
Bachem
Max G
MG
Zhou
Shaoxia
S
Sun
Zilin
Z
Chen
Jinfei
J
Siech
Marco
M
Bimmler
Daniel
D
Graf
Rolf
R
D000951:Antigens, Neoplasm / Q000502:physiology*; D014408:Biomarkers, Tumor / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D058851:GPI-Linked Proteins; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D037181:Lectins, C-Type / Q000502:physiology*; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D010179:Pancreas / Q000166:cytology / Q000187:drug effects*; D000073718:Pancreatitis-Associated Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000037:antagonists & inhibitors* / Q000378:metabolism
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D058851:GPI-Linked Proteins; D037181:Lectins, C-Type; D061965:Matrix Metalloproteinase Inhibitors; D008562:Membrane Glycoproteins; D000073718:Pancreatitis-Associated Proteins; C115460:RECK protein, human; C000612141:REG3A protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1