Polarity of prostate specific membrane antigen, prostate stem cell antigen, and prostate specific antigen in prostate tissue and in a cultured epithelial cell line.
BACKGROUND
Madin-Darby canine kidney (MDCK) cells are immortalized epithelial cells that have been used extensively as a model system to study intracellular molecular trafficking, polarized expression, and secretion of proteins in various epithelia. In order to determine if MDCK cells might serve as a model to study molecular events within prostate epithelial cells, we have evaluated the polarized distribution of three prostate restricted proteins, PSMA, PSCA, and PSA, in situ, and in MDCK cells.


METHODS
Using immunofluorescence, confocal microscopy, cell surface biotinylation, antibody internalization, and biochemical assays we evaluated surface expression and secretion of three prostate restricted proteins expressed in MDCK cells. We compared these patterns of expression to results observed within prostatic epithelium.


RESULTS
We demonstrate that PSMA is localized primarily to the apical plasma membrane in both the prostatic epithelium and transfected MDCK cells, whereas PSCA is expressed in a non-polarized fashion. We also show that PSA is secreted predominantly from the apical surface of transfected MDCK cells, consistent with in vivo observations.


CONCLUSIONS
Similar patterns of localization among MDCK and prostatic epithelial cells suggest that the mechanisms of polarized sorting within these cell types are conserved. Thus, MDCK cells offer a useful model system to study mechanisms of targeting of these proteins within the prostate.
Christiansen
Jason J
JJ
Department of Pathology and Laboratory Medicine, Room 13-344 CHS, University of California Los Angeles, Los Angeles, CA 90095, USA.
Rajasekaran
Sigrid A
SA
Moy
Peggy
P
Butch
Anthony
A
Goodglick
Lee
L
Gu
Zhennan
Z
Reiter
Robert E
RE
Bander
Neil H
NH
Rajasekaran
Ayyappan K
AK
D000818:Animals; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D019921:Biotinylation; D002268:Carboxypeptidases / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D016764:Cell Polarity / Q000502:physiology*; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D058851:GPI-Linked Proteins; D043425:Glutamate Carboxypeptidase II; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000378:metabolism*; D011467:Prostate / Q000378:metabolism*; D017430:Prostate-Specific Antigen / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000378:metabolism*; D014162:Transfection
D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C110843:PSCA protein, human; D002268:Carboxypeptidases; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; D017430:Prostate-Specific Antigen