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Androgen Receptor Stabilization in Recurrent Prostate Cancer Is Associated with Hypersensitivity to Low Androgen¹

Departments of Pediatrics [C. W. G., R. T. J., F. S. F., E. M. W.], Surgery (Division of Urology) [J. L. M.], Pathology and Laboratory Medicine [J. L. M.], and Biochemistry/Biophysics [E. M. W.]; The Laboratories for Reproductive Biology [C. W. G., R. T. J., J. L. M., F. S. F., E. M. W.]; and the Lineberger Comprehensive Cancer Center [C. W. G., J. L. M., F. S. F., E. M. W.], The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599

The androgen receptor (AR) is highly expressed in androgen-dependent and recurrent prostate cancer (CaP) suggesting it has a role in the growth and progression of CaP. Previously proposed mechanisms for AR reactivation in recurrent CaP include altered growth factor signaling leading to protein phosphorylation and AR mutations that broaden ligand specificity. To further establish a role for AR in recurrent CaP, we compared several properties of AR in relation to the growth response to low levels of androgens in model systems of androgen-dependent and recurrent CaP. AR from all of the tumors and cell lines bound [³H]R1881 with similar high affinity (mean K_d, 0.12 nM). In the absence of androgen, AR in androgen-dependent LNCaP cells was unstable with a degradation half-time (t_1/2) of 3 h at 37°C. In contrast, AR was 2–4 times more stable in recurrent CWR22 tumors (t_1/2, >12 h) and CWR-R1 or LNCaP-C4-2 cell lines (t_1/2, 6–7 h) derived from recurrent prostate tumors. In the recurrent CWR22 tumor and its CWR-R1 cell line grown in the absence of androgen, AR immunostaining was entirely nuclear, whereas under the same conditions AR in LNCaP-C4-2 and LNCaP cells was predominantly nuclear but was also detected in the cytoplasm. High level expression, increased stability, and nuclear localization of AR in recurrent tumor cells were associated with an increased sensitivity to the growth-promoting effects of dihydrotestosterone in the femtomolar range. The concentration of dihydrotestosterone required for growth stimulation in CWR-R1 and LNCaP-C4-2 cells was four orders of magnitude lower than that required for androgen-dependent LNCaP cells. The results suggest that AR is transcriptionally active in recurrent CaP and can increase cell proliferation at the low circulating levels of androgen reported in castrated men.

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