Autocrine stimulation of human pancreatic duct-like development by soluble isoforms of epimorphin in vitro.
Epimorphin was recently described as a mesenchymal factor modulating morphogenesis of murine mammary ducts, skin, liver, and lung in vitro. In this study epimorphin was analyzed in a human, pancreatic adenocarcinoma cell line (A818-6) which develops single layer epithelial hollow spheres resembling normal pancreatic ductal structures in vitro. Soluble 34- and 31-kD isoforms of epimorphin were found in the culture supernatant of A818-6 cells. In lysates of A818-6 cells we detected the 34-and 31-kD isoforms and the dimers, and in lysates of fibroblasts the 150-kD tetramers of epimorphin additionally. A neutralizing monoclonal antibody against epimorphin (MC-1) efficiently blocked the development of hollow sphere structures from A818-6 cells. Coculture of A818-6 cells with fibroblasts stimulated the development of hollow sphere structures in general and increased differentiation in 5-6-d-old hollow spheres. A818-6 hollow sphere development in the presence of fibroblasts was also blocked by MC-1. In this novel system for human duct-like differentiation of pancreatic epithelial cells, we provide evidence for an autocrine and paracrine function of epimorphin as a major mediator for morphogenesis.
Molecular Oncology, Department of General and Thoracic Surgery, Christian-Albrechts-University, Kiel, Germany.
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Lehnert
M M
MM
Lerch
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Hirai
M L
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Kruse
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Schmiegel
H
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Kalthoff
D000230:Adenocarcinoma; D000368:Aged; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D019898:Autocrine Communication / Q000187:drug effects; D015415:Biomarkers / Q000032:analysis; D002448:Cell Adhesion; D002449:Cell Aggregation / Q000187:drug effects; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000378:metabolism; D019281:Dimerization; D005260:Female; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D009024:Morphogenesis / Q000187:drug effects; D010183:Pancreatic Ducts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D020033:Protein Isoforms / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D012995:Solubility; D050827:Syntaxin 1; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D017077:Culture Media, Conditioned; D006133:Growth Substances; D008562:Membrane Glycoproteins; D020033:Protein Isoforms; C074558:STX2 protein, human; D050827:Syntaxin 1