Identification of secreted glycoproteins of human prostate and bladder stromal cells by comparative quantitative proteomics.
BACKGROUND
Functional development of the prostate is governed by stromal mesenchyme induction and epithelial response. Stromal/epithelial signaling can be mediated through direct cell-cell contact and diffusible factors and their cell surface receptors. These inducers are likely secreted or membrane-associated extracellular proteins. Given the importance of intercellular communication, it is possible that diseases like cancer could arise from a loss of this communication. One approach to gain a molecular understanding of stromal cells is to identify, as a first step, secreted stromal signaling factors. We proposed to do this by comparative analysis between bladder and prostate.


METHODS
Secreted proteins were identified from cultured normal prostate and bladder stromal mesenchyme cells by glycopeptide-capture method followed by mass spectrometry. Differences in protein abundance between prostate and bladder were quantified from calculated peptide ion current area (PICA) followed by Western validation. Functional and pathway analyses of the proteins were carried out by Gene Ontology (GO) and Teranode software.


RESULTS
This analysis produced a list of 116 prostate and 84 bladder secreted glycoproteins with ProteinProphet probability scores > or =0.9. Stromal proteins upregulated in the prostate include cathepsin L, follistatin-related protein, neuroendocrine convertase, tumor necrosis factor receptor, and others that are known to be involved in signal transduction, extracellular matrix interaction, differentiation and transport.


CONCLUSIONS
We have identified a number of potential proteins for stromal signaling and bladder or prostate differentiation program. The prostate stromal/epithelial signaling may be accomplished through activation of the ECM-receptor interaction, complement and coagulation cascades, focal adhesion and cell adhesion pathways.
Goo
Young Ah
YA
Department of Urology, University of Washington, Seattle, Washington 98195-7610, USA. youngah@u.washington.edu
Liu
Alvin Y
AY
Ryu
Soyoung
S
Shaffer
Scott A
SA
Malmström
Lars
L
Page
Laura
L
Nguyen
Liem T
LT
Doneanu
Catalin E
CE
Goodlett
David R
DR
D015153:Blotting, Western; D002478:Cells, Cultured; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008648:Mesoderm / Q000166:cytology; D009928:Organ Specificity; D011467:Prostate / Q000166:cytology*; D040901:Proteomics; D015398:Signal Transduction / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D001743:Urinary Bladder / Q000166:cytology*
D006023:Glycoproteins
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