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Autocrine effect of DHT on FGF signaling and cell proliferation in LNCaP cells: Role of heparin/heparan‐degrading enzymes
Author(s) -
Kassen Annette E.,
Sensibar Julia A.,
Sintich Sharon M.,
Pruden Shaina J.,
Kozlowski James M.,
Lee Chung
Publication year - 2000
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/1097-0045(20000701)44:2<124::aid-pros5>3.0.co;2-s
Subject(s) - autocrine signalling , lncap , fibroblast growth factor , cell growth , enzyme , heparin , microbiology and biotechnology , signal transduction , cancer research , biology , endocrinology , medicine , cancer , cancer cell , biochemistry , receptor
BACKGROUND LNCaP cells are androgen‐sensitive human prostate cancer cells. They are characterized by a bell‐shaped growth curve in response to increasing doses of dihydrotestosterone (DHT) in culture. At a low concentration of DHT (0.1 nM), these cells show an increase in proliferation, but their growth is arrested at a high concentration (100 nM) of DHT. Results of our previous study demonstrated that the inhibitory effect of DHT at a high concentration was mediated through the action of TGF‐β1. The objective of the present study was to elucidate the mechanism of the proliferative effect of DHT in LNCaP cells. METHODS AND RESULTS DHT stimulated LNCaP proliferation only when cells were cultured in the presence of serum. In serum‐free cultures, the characteristic DHT‐induced proliferation was not observed. The addition of neutralizing antibody against FGF‐2 (basic fibroblast growth factor) was able to inhibit this DHT‐induced proliferation. These results suggest that the proliferative effect of DHT was mediated through the action of FGF‐2. However, results of the reverse transcriptase polymerase chain reaction indicated that LNCaP cells did not express FGF‐2 message. As a result, the source of FGF‐2 in these cultures must be the serum supplemented in the culture media. FGF‐2 can bind to heparin sulfate chains within the extracellular matrix (ECM). In cultures treated with exogenous heparin, the proliferative effect of DHT was abolished. These results led to the development of the hypothesis that DHT treatment mediates the release of FGF‐2 entrapped in the ECM through increased heparinase activity. The addition of heparinase to cultures of LNCaP cells, in the absence of DHT, was able to stimulate cell proliferation. Moreover, 0.1 nM DHT caused a significant increase in heparinase activity. CONCLUSIONS These results provide a possible mechanism for DHT action in LNCaP cells. In the absence of DHT, FGF‐2 in culture was trapped in the extracellular matrix and was not available to interact with LNCaP cells. However, in the presence of 0.1 nM DHT, heparinase activity in the culture was elevated and, as a result, it liberated the trapped FGF‐2 which, in turn, stimulated proliferation in LNCaP cells. Prostate 44:124–132, 2000. © 2000 Wiley‐Liss, Inc.