Different proportions of various prostate-specific antigen (PSA) and human kallikrein 2 (hK2) forms are present in noninduced and androgen-induced LNCaP cells.
BACKGROUND
Human prostate-specific antigen (PSA) and human kallikrein 2 (hK2) are expressed primarily by prostate epithelial cells. PSA and hK2 both exist as free protein and complexed with protease inhibitors (e.g., alpha1-antichymotrypsin, ACT) in serum. The expression of PSA and hK2 in LNCaP cells is upregulated by androgen.


METHODS
LNCaP, a prostate cancer cell line that secretes both PSA and hK2, was used as a model to study the biosynthesis and processing of PSA and hK2 upon androgen induction.


RESULTS
Precursor (zymogen or pro) forms of both PSA and hK2 were detected in spent media of induced and noninduced LNCaP cells, indicating that PSA and hK2 are secreted as proPSA (pPSA) and prohK2 (phK2), respectively, and are converted to the mature forms extracellularly. A 3-fold higher ratio of mature to pPSA was detected in the spent media of mibolerone-induced LNCaP cells compared to noninduced cells. In addition to the inactive proform of PSA, more than half of the mature unclipped PSA present in the spent media did not complex with exogenously added ACT. Spent media of mibolerone-induced LNCaP cells contained nearly 100% mature hK2, whereas the spent media of noninduced cells contained mostly phK2.


CONCLUSIONS
These results indicate that androgens not only upregulate the expression of these kallikreins, but also have a significant effect on the processing of PSA and hK2. These results also show that LNCaP cells express a heterogeneous mixture of inactive PSA and hK2 forms that may serve as a model for the genesis of these forms in physiological fluids. These findings may also provide insights into the forms and ratios of PSA and hK2 in normal and malignant breast tissues.
Kumar
A
A
Hybritech Incorporated, a subsidiary of Beckman Coulter, Inc., San Diego, California 92196-9006, USA.
Mikolajczyk
S D
SD
Hill
T M
TM
Millar
L S
LS
Saedi
M S
MS
D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D009277:Nandrolone / Q000031:analogs & derivatives* / Q000494:pharmacology; D011467:Prostate / Q000201:enzymology; D017430:Prostate-Specific Antigen / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D020033:Protein Isoforms; D020539:Sequence Analysis, Protein; D045165:Testosterone Congeners / Q000494:pharmacology*; D020840:Tissue Kallikreins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D014407:Tumor Cells, Cultured; D000514:alpha 1-Antichymotrypsin / Q000737:chemistry
D000911:Antibodies, Monoclonal; D020033:Protein Isoforms; D045165:Testosterone Congeners; D000514:alpha 1-Antichymotrypsin; D009277:Nandrolone; C100075:mibolerone; D020840:Tissue Kallikreins; D017430:Prostate-Specific Antigen