Effects of alpha(1)-adrenoceptor (alpha(1)-AR) antagonists on cell proliferation and apoptosis in the prostate: therapeutic implications in prostatic disease.

BACKGROUND: Benign prostate hyperplasia (BPH) and prostate cancer established that disruption of the molecular mechanisms that regulate apoptosis and cell proliferation among the stromal and epithelial cell populations, may underlie the neoplastic development that characterizes the aging gland. This work examined the effects of selected alpha(1)-adrenoceptor (alpha(1)-AR) antagonists (blockers) on cellular dynamics to determine whether induction of apoptosis or inhibition of proliferation could contribute to the overall clinical profile. METHODS: Our efforts were focused on investigating whether alpha(1)-AR antagonists of two different chemical classes affect prostate pathophysiology via mechanisms other than smooth muscle contraction. In in vitro experiments, the two clinically used quinazoline alpha(1)-adrenoceptor antagonists terazosin and doxazosin and the chemically-distinct sulphonamide, tamsulosin, were examined for effects on prostatic tumor growth, by inhibiting cell proliferation and'or inducing apoptosis. RESULTS: Our findings suggest that alpha(1)-AR antagonists, terazosin and doxazosin, suppress prostatic growth by inducing apoptosis in a dose-dependent manner and without affecting cell proliferation. Tamsulosin exerted no effect on prostate cancer cell growth. The apoptotic effect of terazosin and doxazosin appears to be independent of the alpha(1)-adrenoceptor block. CONCLUSIONS: Taken together, our findings demonstrate the ability of the quinazoline alpha-blockers, terazosin and doxazosin, but not the sulphonamide, tamsulosin, to suppress prostate growth by inducing apoptosis among the epithelial cells in the benign and malignant prostate. These studies underwrite the durability of the response seen in long-term studies with terazosin, and suggest the potential of this drug in the treatment of prostate carcinoma.

Suppression of human prostate cancer cell growth by alpha1-adrenoceptor antagonists doxazosin and terazosin via induction of apoptosis.

Recent evidence from our laboratory has demonstrated that alpha1-adrenoceptor antagonists doxazosin and terazosin induced apoptosis in prostate epithelial and smooth muscle cells in patients with benign prostatic hypertrophy (BPH; J. Urol., 159: 1810-1815, 1998; J. Urol., 161: 2002-2007, 1999). In this study, we investigated the biological action of three alpha1-adrenoceptor antagonists, doxazosin, terazosin, and tamsulosin, against prostate cancer cell growth. The antigrowth effect of the three alpha1-adrenoceptor antagonists was examined in two human prostate cancer cell lines, PC-3 and DU-145, and a prostate smooth muscle cell primary culture, SMC-1, on the basis of: (a) cell viability assay; (b) rate of DNA synthesis; and (c) induction of apoptosis. Our results indicate that treatment of prostate cancer cells with doxazosin or terazosin results in a significant loss of cell viability, via induction of apoptosis in a dose-dependent manner, whereas tamsulosin had no effect on prostate cell growth. Neither doxazosin nor terazosin exerted a significant effect on the rate of cell proliferation in prostate cancer cells. Exposure to phenoxybenzamine, an irreversible inhibitor of alpha1-adrenoceptors, does not abrogate the apoptotic effect of doxazosin or terazosin against human prostate cancer or smooth muscle cells. This suggests that the apoptotic activity of doxazosin and terazosin against prostate cells is independent of their capacity to antagonize alpha1-adrenoceptors. Furthermore, an in vivo efficacy trial demonstrated that doxazosin administration (at tolerated pharmacologically relevant doses) in SCID mice bearing PC-3 prostate cancer xenografts resulted in a significant inhibition of tumor growth. These findings demonstrate the ability of doxazosin and terazosin (but not tamsulosin) to suppress prostate cancer cell growth in vitro and in vivo by inducing apoptosis without affecting cell proliferation. This evidence provides the rationale for targeting both drugs, already in clinical use and with established adverse-effect profiles, against prostatic tumors for the treatment of advanced prostate cancer.

Overexpression of human epidermal growth factor receptor and c-erbB-2 by neuroendocrine cells in normal prostatic tissue.

OBJECTIVE: It has been suggested that prostatic neuroendocrine (NE) cells play an important role in the growth and differentiation of the prostate by secreting various neuropeptides and serotonin. However, the mechanism by which NE cells themselves are regulated is virtually unknown. In the present study we evaluated the expression of the human epidermal growth factor receptor (EGFR) family (HER) in prostatic NE cells. METHODS: Formalin-fixed, paraffin-embedded tissue sections from twenty radical prostatectomy specimens were immunostained with validated rabbit polyclonal antibodies raised against human EGFR and c-erbB-2, using the streptavidin-peroxidase enzyme conjugate method. RESULTS: A strong immunoreactivity was observed with both antibodies in the cytosol of a few epithelial cells. These cells frequently had a dendritic appearance and were located in the acini and ducts. The EGFR-positive cells were predominant in most cases. Double immunostaining revealed the colocalization of both antigens with chromogranin A, a polypeptide that is expressed by most NE cells. Moreover, EGFR and c-erbB-2 appeared to be colocalized as well as independently expressed by different subpopulations of NE cells. CONCLUSIONS: The results suggest that prostatic NE cells might be regulated by the HER protein family, probably, in a ligand-specific fashion. This is the first report identifying a potential pathway regulating prostatic NE cells.

Androgen receptor immunostaining and its tissue distribution in formalin-fixed, paraffin-embedded sections after microwave treatment.

We used microwave (MW) oven heat treatment to unmask human androgen receptor (AR) immunostaining in formalin-fixed, paraffin-embedded tissue. Prostate tissue was used as an AR-positive control. Tissue sections were boiled in citrate buffer in a conventional MW oven for 30 min, followed by immunostaining with a validated murine monoclonal antibody (MAb), F39.4.1, raised against a peptide included in the N-terminal domain of the 100 KD human AR. AR immunostaining was localized to the nuclei of prostate secretory epithelial cells but was weak or absent in basal cells and of variable intensity in the stromal cells. Slides exposed to less than 10 min of MW heat treatment or none at all manifested no AR immunoreactivity. Tissue morphology was well preserved. Immunohistochemical determination of AR status in a wide variety of human tissues was consistent with that previously reported by others using frozen sections. MW heat treatment of tissue samples in an excellent method of localizing AR antigenicity, enabling immunohistochemical evaluation of AR status in formalin-fixed, paraffin-embedded material.

Immunohistochemical localization of parathyroid hormone-related protein in human prostate cancer.

Parathyroid hormone-related protein (PTHrP) is produced by a variety of malignant tumors and has been implicated as a major cause of humoral hypercalcemia of malignancy. Expression of PTHrP in prostate cancer tissue was studied immunohistochemically using 33 radical prostatectomy specimens from patients with clinically localized carcinoma of the prostate. None of these patients demonstrated hypercalcemia prior to the surgery. Acetone-methyl benzoate-xylene-processed, paraffin-embedded tissues were stained with a validated mouse monoclonal antibody to an amino acid fragment, PTHrP(109-141), using the streptavidin-peroxidase enzyme conjugate method. All cases (33 of 33; 100%) studied demonstrated some degree of immunoreactivity throughout the cytoplasm of the tumor cells, but immunostaining was absent from inflammatory and stromal cells. The intensity of the staining appeared to directly correlate with increasing tumor grade. The widespread immunohistochemical localization of PTHrP in carcinoma of the prostate suggests that PTHrP may play some local role in the growth of transformed cells in the prostate. Furthermore, overexpression of PTHrP may be a possible marker to evaluate the malignant potential of carcinoma of the prostate.