The neuroendocrine phenotype of gastric myofibroblasts and its loss with cancer progression.
Stromal cells influence cancer progression. Myofibroblasts are an important stromal cell type, which influence the tumour microenvironment by release of extracellular matrix (ECM) proteins, proteases, cytokines and chemokines. The mechanisms of secretion are poorly understood. Here, we describe the secretion of marker proteins in gastric cancer and control myofibroblasts in response to insulin-like growth factor (IGF) stimulation and, using functional genomic approaches, we identify proteins influencing the secretory response. IGF rapidly increased myofibroblast secretion of an ECM protein, TGFβig-h3. The secretory response was not blocked by inhibition of protein synthesis and was partially mediated by increased intracellular calcium (Ca(2+)). The capacity for evoked secretion was associated with the presence of dense-core secretory vesicles and was lost in cells from patients with advanced gastric cancer. In cells responding to IGF-II, the expression of neuroendocrine marker proteins, including secretogranin-II and proenkephalin, was identified by gene array and LC-MS/MS respectively, and verified experimentally. The expression of proenkephalin was decreased in cancers from patients with advanced disease. Inhibition of secretogranin-II expression decreased the secretory response to IGF, and its over-expression recovered the secretory response consistent with a role in secretory vesicle biogenesis. We conclude that normal and some gastric cancer myofibroblasts have a neuroendocrine-like phenotype characterized by Ca(2+)-dependent regulated secretion, dense-core secretory vesicles and expression of neuroendocrine marker proteins; loss of the phenotype is associated with advanced cancer. A failure to regulate myofibroblast protein secretion may contribute to cancer progression.
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Silvia
S
Balabanova
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Chris
C
Holmberg
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Islay
I
Steele
Institute of Integrative Biology, University of Liverpool, Liverpool, L69 3BX UK.
Bahram
B
Ebrahimi
Institute of Integrative Biology, University of Liverpool, Liverpool, L69 3BX UK.
Lucille
L
Rainbow
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Ted
T
Burdyga
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Cathy
C
McCaig
Department of Pathology and.
Lazso
L
Tiszlavicz
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Nantaporn
N
Lertkowit
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Olivier T
OT
Giger
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Simon
S
Oliver
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Ian
I
Prior
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Rod
R
Dimaline
Institute of Integrative Biology, University of Liverpool, Liverpool, L69 3BX UK.
Deborah
D
Simpson
Institute of Integrative Biology, University of Liverpool, Liverpool, L69 3BX UK.
Rob
R
Beynon
Department of Medicine, University of Szeged, Szeged, H-6701 Hungary.
Peter
P
Hegyi
Department of Medicine, Columbia University, New York, NY 10032-3802, USA and.
Timothy C
TC
Wang
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK.
Graham J
GJ
Dockray
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK, Department of Molecular and Clinical Cancer, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX UK avarro@liv.ac.uk.
Andrea
A
Varro
D015153:Blotting, Western; D016022:Case-Control Studies; D002478:Cells, Cultured; D018450:Disease Progression; D005089:Exocytosis / Q000502:physiology; D005753:Gastric Mucosa / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D007335:Insulin-Like Growth Factor II / Q000378:metabolism*; D007553:Isotope Labeling; D058628:Myofibroblasts / Q000378:metabolism / Q000473:pathology*; D009490:Neurosecretory Systems / Q000378:metabolism / Q000473:pathology*; D010641:Phenotype; D034741:RNA, Small Interfering / Q000235:genetics; D053381:Secretogranin II / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000378:metabolism / Q000473:pathology*; D053719:Tandem Mass Spectrometry
D034741:RNA, Small Interfering; C506135:SCG2 protein, human; D053381:Secretogranin II; D007335:Insulin-Like Growth Factor II