Biomedical subjects

H Klocker

Publications and source records attributed to H Klocker.

At least 91 records · Page 5Linked to original sources

PubMed1997

Hyperactive androgen receptor in prostate cancer: what does it mean for new therapy concepts?

Investigations on androgen signaling alterations in the late stages of prostate cancer revealed new molecular mechanisms that may be in part responsible for failure of endocrine therapy. Both primary and metastatic lesions from prostate cancer express androgen receptor protein. Amplification of androgen receptor gene occurs in a subset of prostate cancer patients. Several point mutations of androgen receptor gene have been described; they generate receptors which are functionally activated by androgens, other steroids, and even by antihormones. The frequency of androgen receptor mutations may be high in tumor metastases. Functional activity of androgen receptor is influenced by nonsteroidal factors, such as peptide growth factors and second messengers. Thus, prostate cancer cells adapt to low androgen environment by various mechanisms utilizing androgen receptor. Therefore, new strategies for switching off the androgen receptor are needed.

Animals↗

PubMed1997

Androgen receptor point mutations as the underlying molecular defect in 2 patients with androgen insensitivity syndrome.

PURPOSE: We have detected androgen receptor (AR) gene mutations as the underlying molecular defect in male pseudohermaphroditism patients. The functional properties of these mutant receptors regarding hormone binding and transactivation were characterized in 2 patients and offered a possible treatment modality for a male pseudohermaphroditism newborn. MATERIALS AND METHODS: Specific binding of dihydrotestosterone, thermostability of the receptor-hormone complex and 5alpha-reductase activity were measured in their genital skin fibroblasts. AR gene mutations were detected by direct sequencing. The ability of the mutant receptors to activate androgen responsive elements in the deoxyribonucleic acid was determined by transactivation experiments. Screening techniques that distinguish between normal and mutant ARs were developed for rapid detection of the mutation in other family members. RESULTS: The 2 patients showed a qualitative and quantitative binding defect. In both patients, point mutations in the ligand binding domain were identified as the underlying cause. Transactivation assays demonstrated that in the newborn increasing androgen concentrations can restore the mutated receptor's function completely, whereas in the patient with complete androgen insensitivity, excessive amounts of synthetic androgens were necessary. Therefore, the newborn received androgen stimulation and underwent surgical correction in the male direction. These experiments revealed that the functional difference between a mutant AR that causes a partial and one that causes a complete androgen insensitivity may be very small. CONCLUSIONS: Identification of the molecular mechanisms that cause the various forms of sex ambiguity will greatly improve both diagnosis and therapy in affected patients. Exact characterization of AR activation and function may offer a possible treatment modality in affected patients.

Child↗

PubMed1996

Expression of the gene for tear lipocalin/von Ebner's gland protein in human prostate.

Northern analysis of human multiple tissue blots containing poly A+ RNA from spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes revealed that a prostate specific transcript hybridizes to a tear lipocalin/von Ebner's gland protein (TL/VEGP) gene probe. To characterize this transcript, the corresponding cDNA was amplified by reverse transcription (RT)-PCR. Cloning and sequence analysis showed that it was identical to the tear lipocalin cDNA isolated from human lachrymal glands. Immunohistochemical analysis on thin layer sections of human prostate using a tear lipocalin specific antiserum confirmed the expression of this cDNA in prostate. Thus, our results clearly argue against a unique function of TL/VEGP in human tear fluid or saliva. The human cDNA was expressed in E. coli using the pQE system yielding a recombinant protein which shows biochemical properties identical to the native TL/VEGP.

Amino Acid Sequence↗

PubMed1996

Human renal-cell carcinoma tissue contains dendritic cells.

Immune surveillance of cancer requires antigen-presenting cells which activate T cells specific for tumor-associated antigens. We show here that substantial numbers of dendritic cells, which are the most potent antigen-presenting cells, emigrate from renal-tumor explants in organ culture. Tumor-derived dendritic cells presented with all characteristics of mature dendritic cells. Dendritic cells could be identified by typical cytoplasmic projections (=veils). They expressed high levels of MHC products and of the co-stimulator CD86 (B7-2). Dendritic cells expressed the CD45RO isoform but not CD45RA. The most important point was that up to 9% of the emigrating leukocytes expressed the CD83 antigen, a specific marker for mature dendritic cells. CD83+ cells were approximately 40-fold enriched in the tumor tissue as compared to the peripheral blood. In contrast to cultured blood dendritic cells, tumor-emigrant dendritic cells had a reduced potential to capture soluble antigen, as shown by the exclusion of fluoresceinated Dextran molecules. Finally, in mixed leukocyte reactions, tumor-derived dendritic cells were able to stimulate naive T cells from cord blood, which is a unique feature of dendritic cells. This study demonstrates that genuine dendritic cells reside in or infiltrate renal-cell carcinoma tissue. The failure of patients with renal-cell carcinoma to mount an anti-tumor immune response despite the presence of professional antigen-presenting cells in the tumor tissue suggests that tumor-associated dendritic cells are suppressed in situ, in a similar way to that described for tumor-infiltrating lymphocytes.

Antigens, CD↗

PubMed1996

Androgen receptor-Ets protein interaction is a novel mechanism for steroid hormone-mediated down-modulation of matrix metalloproteinase expression.

Matrix metalloproteinases belong to a family of structurally related enzymes that plays important role in tissue morphogenesis, differentiation, and wound healing. Their expression is negatively regulated by several members of the steroid hormone receptor family. This is thought to occur through interaction of the steroid receptors with the transcription factor AP-1 that is otherwise required for positive regulation. Here, we demonstrate that AP-1 is not always a target for down-regulation of expression of matrix metalloproteinases by steroid receptors. Androgen receptor negatively regulates matrix metalloproteinase-1 expression not through AP-1 but through a family of Ets-related transcription factors that are also required for positive regulation. This negative regulation is specific for the androgen receptor. It does not require the DNA binding activity but needs amino-terminal sequences of the receptor. These results identify a novel regulatory pathway for negative regulation utilized by a member of the steroid hormone receptor family for down-regulating the expression of matrix metalloproteinases.

Androgens↗

PubMed1996

Evaluation and comparison of two new prostate carcinoma markers. Free-prostate specific antigen and prostate specific membrane antigen.

BACKGROUND: Two new prostate cancer markers, free-prostate specific antigen (f-PSA) and prostate specific membrane antigen (PSMA) were recently introduced. This report summarizes a prospective two-year multicenter test of their diagnostic or prognostic capabilities. Total PSA was also measured. METHODS: There were four clinical groups studied: (1) 226 individuals from a screening project undergoing ultrasound and biopsy evaluation had markers obtained: (2) 68 patients suspected of having prostate cancer and undergoing 2 or more biopsies had the markers obtained on multiple occasions: (3) 100 patients undergoing radical prostatectomy had markers obtained pre- and post-operatively: and (4) 31 patients with metastatic prostate cancer each had multiple samples for marker assay obtained over a 2-year period. In all, 465 patients had one or more samples obtained and studied. RESULTS: Free-PSA affords little additional diagnostic advantage compared with total PSA in the screening population. The receiver operating characteristic curves for diagnostic accuracy were ranked: (1) PSA density; (2) total PSA; (3) f-PSA; and (4) PSMA, PSMA showed the best correlation with stage of the primary tumor in the screened group. In the multiple negative biopsy group, f-PSA varied from 12 to 21%. PSMA values were evaluated in all histologic categories. PSA density was > or = 0.15 in all categories. In the prostatectomy cases PSA values postoperatively were quite low in Stage II; f-PSA was of no value. Later, f-PSA was increased in association with elevated total PSA values. Mean PSMA values were above normal in all postoperative time periods except in Stage III patients at 6 months to 1 year postoperatively. PSA densities were all > or = 0.15. In patients with metastatic carcinoma, elevated PSMA values correlated best with a poor prognosis (clinical progression), as has been described. CONCLUSIONS: These data suggest that f-PSA values do not provide additional diagnostic benefit compared with total PSA in screening populations, in the presence of suspected cancer, postprostatectomy, or in metastatic disease. PSMA is of prognostic significance, especially in the presence of metastatic disease, and correlates well with the stage of disease in cancers detected in a screened population.

Antigens, Neoplasm↗

PubMed1996

Characterization of detergent-solubilized beef liver mitochondrial NAD+ glycohydrolase and its truncated hydrosoluble form.

Membrane-bound beef liver mitochondrial NAD+ glycohydrolase (NADase) was partially purified after its solubilization by either detergent or crude pancreatic lipase, steapsin. Solubilization by steapsin yielded a homogeneous water-soluble enzyme. A fluorescence assay was developed that allowed visualization of NADase activity directly within the gel after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The apparent molecular masses of the detergent- and steapsin-solubilized forms were estimated to be about 30,000 and 28,000, respectively. The small part that was cleaved by steapsin represents presumably the membrane anchor of the mitochondrial NADase, as its removal converted the enzyme from a highly hydrophobic to a hydrosoluble protein. The fluorescence staining for activity was also successfully applied to other NADases. Kinetic analyses of the two forms of solubilized mitochondrial NADase revealed that the catalytic properties were unaffected after the steapsin treatment. Neither the binding affinity of the substrate analog 1, N6-etheno-NAD+ nor the inhibition by nicotinamide differed significantly between these two forms of the enzyme. Moreover, the dependence of the enzyme activity on temperature, pH, or ionic strength was also similar for both preparations. However, activity of the detergent-solubilized but not of the truncated steapsin-solubilized enzyme was strongly dependent on the presence of bivalent metal ions such as ZN2+. These results suggest that the membrane part of the mitochondrial NAD+ glycohydrolase is not required for catalysis. It appears, however, to be of importance for the regulation of the enzyme.

Animals↗

PubMed1996

CAG-repeat expansion in androgen receptor in Kennedy's disease is not a loss of function mutation.

Expansion of CAG trinucleotide repeats in androgen receptor gene is present in patients with a rare X-linked inherited form of motor neuron disorder termed Kennedy's disease or spinal and bulbar muscular atrophy (SBMA). This is a late onset progressive disease often associated with mild signs of androgen insensitivity. Defects in androgen receptor (AR) action have been linked to the expansion of the CAG trinucleotide repeats and postulated to be the cause of the disease. We have identified a trinucleotide repeat of 45 in the N-terminus of the AR in two brothers with SBMA and several members in their family (range in the general population is 11-35). Treatment of the patients with androgens failed to improve their clinical symptoms and provided no hint of an anomalous function of the AR. Consistently, functional analysis of the mutant receptor showed hormone binding, transactivation and transrepression potentials identical to that of the wild-type receptor. These results together argue against SBMA being a loss of function mutation of the AR.

Androgens↗

PubMed1996

Androgen receptor status of lymph node metastases from prostate cancer.

To date androgen receptor (AR) expression and structure in human prostatic cancer have been studied in primary tumor specimens and in cell lines. Investigation of alterations in the androgen-signalling transduction cascade in prostatic carcinoma metastases is important to improve our understanding of tumor progression towards androgen insensitivity. In the present study we have collected data comparing AR expression in both the primary tumors and the respective pelvic lymph node metastases. Formalin-fixed and paraffin-embedded tissues derived from the primary tumors and positive lymph nodes of 12 patients undergoing radical prostatectomy were immunostained for the AR and prostate-specific antigen (PSA). AR expression was evaluated with the polyclonal antibody PG-21, which is directed against amino acid 1-21 in the N-terminal region of the AR. All primary tumors stained for the AR. In 8 of the 12 lymph nodes examined more than 50% of the tumor cells were AR positive and displayed a uniform staining pattern; in one lymph node metastasis remarkable heterogeneity in AR expression was observed. In two cases less than 10% of the tumor cells stained for the AR. In one case the lymph node metastasis was immunohistochemically negative for the AR, whereas the primary tumor obtained from the same patient displayed intense staining for the AR. PSA was expressed in all metastases and primary tumors. Our data demonstrate that loss of the AR in lymph node metastases from prostatic carcinoma is a rare event.

Adenocarcinoma↗

PubMed1996

Inhibitory effects of the nucleoside analogue gemcitabine on prostatic carcinoma cells.

Gemcitabine (2',2'difluoro-2'deoxycytidine, dFdC) is a synthetic antimetabolite of the cellular pyrimidine nucleotide metabolism. In a first series of in vitro experiments, the drug showed a strong effect on the proliferation and colony formation of the human androgen-sensitive tumor cell line LNCaP and the androgen-insensitive cell lines PC-3 and DU-145. Maximal inhibition occurred at a dFdC concentration as low as 30 nM. In contrast to the cell lines which were derived from metastatic lesions of prostate cancer patients, no inhibitory effects were found in normal primary prostatic epithelial cells at concentrations up to 100 nM. The effect of gemcitabine was reversed by co-administration of 10-100 microM of its natural analogue deoxycytidine. In view of a future clinical application of this anti-tumor drug in advanced prostatic carcinoma, we have compared the effect of gemcitabine on prostatic tumor cells with that on bone marrow granulopoietic-macrophage progenitor cells, because neutropenia is a common side effect of gemcitabine treatment. The time course of action on the two kinds of cells was markedly different. Colony formation of tumor cells was inhibited by two thirds at a gemcitabine concentration of about 3.5 nM. The same effect on granulopoietic-macrophagic progenitor cells required a concentration of 9 nM. Co-administration of deoxycytidine to gemcitabine-treated tumor cell cultures completely antagonized the effect of gemcitabine whereas addition of deoxycytidine after 48 hr of gemcitabine treatment could not prevent gemcitabine action on the tumor cells. In contrast, more than half of the granulopoietic-macrophagic progenitor cells could still be rescued by deoxycytidine administration after 48 hr. These findings and the marked difference in the susceptibility of neoplastic and normal prostatic cells suggest that gemcitabine is a promising substance which should be further evaluated as to its efficacy in the treatment of advanced prostatic carcinoma.

Antimetabolites, Antineoplastic↗

PubMed1996

Regulation of prostatic growth and function by peptide growth factors.

Polypeptide growth factors are positive and negative regulators of prostatic growth and function. Expression and biological effects of epidermal growth factor (EGF), transforming growth factors (TGFs) alpha and beta, fibroblast growth factors (FGFs), and insulin-like growth factors (IGFs) in the prostate have been extensively studied. EGF and TGF alpha, which share the same receptor, are strong mitogens for prostatic epithelial and stromal cells. Their paracrine mode of action in normal tissue and early-stage tumors is apparently altered towards an autocrine stimulation in hormone-independent tumors, which gain the ability to produce TGF alpha by themselves. TGF beta has a dual role in the regulation of prostatic growth. It inhibits growth of prostatic epithelial cells in culture and mediates programmed cell death after androgen withdrawal. However, advanced prostatic carcinomas become insensitive to the inhibitory effect of TGF beta. Several members of the FGF family have been identified in the prostate. They are mainly or exclusively expressed in the stromal cells, and stimulate the epithelial cells. In the rat Dunning tumor model, progression is accompanied by distinct changes in the expression of FGFs and their receptors. In the hyperplastic tissue, basic FGF (bFGF) is accumulated. This growth factor is also a potent angiogenic inducer, expression of which may determine the metastatic capability of a tumor. IGFs are paracrine growth stimulators in the normal and hyperplastic prostate. It is still under consideration whether prostatic cancer cells gain the ability to produce IGF-I by themselves and thus shift to an autocrine mode of IGF-I stimulation. Growth factors also interact with the androgen-signaling pathway. IGF-I in particular, other growth factors as well, can activate the androgen receptor.

Animals↗

PubMed1996

Usefulness of the ratio free/total prostate-specific antigen in addition to total PSA levels in prostate cancer screening.

OBJECTIVES: Two different studies were performed. The aim of the first study was to define whether the measurement of the ratio between free and total prostate-specific antigen (f/t PSA) in serum may enhance the ability of PSA-based screening for early detection of prostate cancer in men with elevated serum PSA levels. A second study was undertaken to investigate the value of f/t PSA ratio in serum to improve the specificity of prostate cancer screening in men with serum PSA levels between 2.5 and 10.0 ng/mL. METHODS: In a retrospective study of 266 men with elevated PSA levels and proven biopsy results, f/t PSA levels were measured using deep frozen serum samples. In a second study we enrolled 158 men with elevated PSA levels according to age reference ranges apparent from our current PSA screening study with additional measurement of the f/t PSA ratio. All study volunteers with a free f/t PSA ratio cutoff point of < or = 22% underwent digital rectal examination, transrectal ultrasonography, and biopsy of the prostate. Free and total PSA levels were measured with the Delfia PSA dual label f/t PSA kit (Wallac Oy Turku, Finland). RESULTS: 106 of 158 men with elevated total PSA values between 2.5 and 10.0 ng/mL (group 1) have been further evaluated and 37 prostate cancers were detected. Mean percentage of free PSA was 10% in men with cancer and 22% in men with benign prostatic hyperplasia. Using a f/t PSA ratio of < or = 22% as a biopsy criterion 30% of the negative biopsies could be eliminated while still detecting 98% carcinomas. CONCLUSIONS: Measurement of f/t PSA reduces the number of unnecessary biopsies in PSA screening without missing many cancers.

Aged↗

PubMed1996

Electromotive diffusion (EMD) and photodynamic therapy with delta-aminolaevulinic acid (delta-ALA) for superficial bladder cancer.

Recent reports have shown that topical application of delta-aminolaevulinic acid (delta-ALA) can be used for the photodynamic diagnosis and therapy of superficial bladder tumours. Electromotive diffusion (EMD) increases the cellular uptake of polar substances. In six patients with biopsy-proven recurrent carcinoma in situ of the bladder after Bacillus Calmette-Guérin (BCG) treatment, 100 cm3 of a 0.5% delta-ALA solution at pH approximately 6 was instilled into the bladder. With a special Foley catheter containing a stainless steel electrode, EMD, with a 15 mA, pulsed current, positive polarity at the catheter, was created for 20 min in the sedated patient. Thereafter a laser fibre with a spheric diffuser was inserted into the bladder over a cystoscope. The bladder surface was irradiated at a wavelength of 632 nm with 350 mW s-1 (total dose, 30-50 J cm-2). Follow-up consisted of a cystoscopy and biopsy after 6 weeks and cystoscopy and cytology every 3 months thereafter. The whole procedure was well tolerated. Currently, five patients are tumour free after a follow-up of 10-16 months. One patient has recurred after 10 months with a Ta G3 superficial tumour at the bladder neck which was resected. In a transwell cell culture model, we investigated the effect of delta-ALA treatment with and without application of EMD. In this in vitro system, application of an electric field showed only a small increase in delta-ALA uptake compared with uptake by passive diffusion.

Aged↗

PubMed1996

Transforming growth factors-beta 1 and beta 2 in serum and urine from patients with bladder carcinoma.

PURPOSE: Transforming growth factors-beta (TGF-beta) are cellular regulators and potent angiogenic factors. We determined serum and urinary levels of TGF-beta 1 and beta 2 in patients with bladder carcinoma to study a correlation with tumor stage, grade and metastatic spread. MATERIALS AND METHODS: Using commercial immunoassays TGF-beta 1 and beta 2 were determined in serum and urine samples from 57 bladder cancer patients and 18 healthy controls. RESULTS: Serum TGF-beta 1 levels were significantly elevated in 21 patients with invasive bladder cancer (61.5 ng./ml.) compared to 18 healthy controls (36.3 ng./ml.), whereas serum TGF-beta 1 levels in 36 patients with superficial bladder tumors were within the normal range (33.4 ng./ml.). Serum TGF-beta 1 was increased in 27 patients with grade 3 tumors (55.7 ng./ml.), compared to 16 with grade 1 and 14 with grade 2 tumors (32.6 and 33.3 ng./ml., respectively). By contrast, serum TGF-beta 2 levels were not different from those of controls. No significant increase in serum TGF-beta 1 and beta 2 could be found in patients with lymph node metastases. In urine specimens there was no significant correlation of TGF-beta 1 and beta 2 with tumor stage. CONCLUSIONS: Our results suggest that elevated serum TGF-beta 1 may be relevant for diagnosis of bladder cancer and further studies are warranted.

Adult↗

PubMed1996

Activation of two mutant androgen receptors from human prostatic carcinoma by adrenal androgens and metabolic derivatives of testosterone.

The androgen receptor (AR) plays a central regulatory role in prostatic carcinoma and is a target of androgen ablation therapy. Recent detection of mutant receptors in tumor specimens suggest a contribution of AR alterations to progression towards androgen independence. In a specimen derived from metastatic prostate cancer we have reported a point mutation in the AR gene that leads to a single amino acid exchange in the ligand binding domain of the receptor. Another amino acid exchange resulting from a point mutation was also identified 15 amino acids away from our mutation. This mutation was detected in the AR gene isolated from an organ-confined prostatic tumor. Here we report the functional characterization of the two mutant receptors in the presence of adrenal androgens and testosterone metabolites. These studies were performed by cotransfecting androgen-responsive reporter genes and either the wild-type or mutant AR expression vectors into receptor negative DU-145 and CV-1 cells. The indicator genes used consisted of the promoter of the androgen-inducible prostate-specific antigen gene or the C' Delta9 enhancer fragment from the promoter of the mouse sex-limited protein driving the expression of the bacterial chloramphenicol acetyl transferase gene. Cotransfection-transactivation assays revealed that the adrenal androgen androstenedione and two products of testosterone metabolism, androsterone and androstandiol, induced reporter gene activity more efficiently in the presence of the mutant receptors than in the presence of the wild-type receptor. No difference between wild-type and mutant receptors was observed in the presence of the metabolite androstandione. The interaction of receptor-hormone complexes with target DNA was studied in vitro by electrophoretic mobility shift assays (EMSA). Dihydrotestosterone and the synthetic androgen mibolerone induced a faster migrating complex with all receptors, whereas the androgen metabolite androstandione induced this complex only with the two mutant receptors. Androsterone and androstandiol were inactive in the EMSA. These aberrant properties of the mutant receptors in the presence of adrenal androgens and products of androgen metabolism may be of importance in the course of the prostate cancer, especially during androgen ablation therapy.

Androgens↗

PubMed1995

Identification and purification of a bovine liver mitochondrial NAD(+)-glycohydrolase.

Nonenzymatic ADP-ribosylation of mitochondrial proteins is thought to play a role in the regulation of Ca2+ efflux from mitochondria. It has been shown that intramitochondrial ADP-ribose is generated by a specific NAD(+)glycohydrolase, which catalizes hydrolysis of NAD+ to ADP-ribose and nicotinamide. We purified this enzyme from bovine liver mitochondrial membranes. The final preparation had a 1660-fold purified enzyme activity and contained a main protein band with an apparent molar mass of 32,000 in a SDS-polyacrylamide gel. The identity of this protein band with NAD(+)-glycohydrolase was verified by renaturation of its enzymatic activity. Partial amino acid sequence information was obtained from two enzyme fragments after proteolytic cleavage of the protein band in the SDS-polyacrylamide gel. Searches in protein databases revealed that an arginine ADP-ribosyl hydrolase harbours two stretches of amino acids that are highly similar to the partial NAD(+)-glycohydrolase sequences.

Amino Acid Sequence↗

PubMed1995

Dendritic antigen-presenting cells from the peripheral blood of renal-cell-carcinoma patients.

Dendritic cells are considered to be the initiators of immune responses, including those directed against tumors. Clinical research on dendritic cells was long hampered by the limited availability of these cells. The recent identification of cytokine combinations that mobilize dendritic cells with potent antigen-presenting cell function from peripheral blood represented a major progress. We show in this study that substantial numbers of dendritic cells can be obtained from the peripheral blood of patients with renal-cell carcinoma. The procedure requires a relatively small blood sample (40 ml) and avoids both priming of the patient with granulocyte-colony stimulating factor and leukapheresis. Approximately 2 to 8 million cells with the characteristics of dendritic cells could be obtained: phase-contrast microscopy revealed the typical cytoplasmic processes or veils; phenotypic analysis confirmed expression of dendritic-cell-associated molecules, including MHC class II, CD1a, CD4, ICAM-1 (CD54), LFA-3 (CD58), B7-1 (CD80) and B7-2 (CD86), and absence of T-cell, B-cell and monocyte markers; in addition, these cells rapidly attached to and migrated on collagen-type-1-coated surfaces. Interestingly, attachment was accompanied by acquisition of the CD14 antigen; functionally, cultured dendritic cells proved to be very potent co-stimulators of the phytohemagglutinin-induced proliferation of autologous tumor-infiltrating lymphocytes. The reproducible growth of functional dendritic cells from cancer patients is encouraging for the design of immunotherapy protocols.

Adult↗

PubMed1995

Mutant androgen receptors in prostatic tumors distinguish between amino-acid-sequence requirements for transactivation and ligand binding.

Mutant androgen receptors are thought to contribute to hormone resistance in prostate carcinoma. The part they play in this process, however, is ill-defined. Here we report on transactivation by 2 mutant androgen receptors from prostatic tumors with single amino-acid exchanges in their hormone-binding domains. These exchanges enhance the transactivation property of the receptors, particularly to androsterone and androstanediol, 2 metabolized derivatives of testosterone present in the prostate. Additionally, they enhance the transactivation potential of the mutant receptors to hydroxyflutamide, an anti-androgen frequently used in hormone ablation therapy. The increased transactivation by the mutant receptors did not result from altered affinity of the receptors to the inducing ligands nor from measurable changes in conformation of the liganded receptors. Thus the single amino-acid exchanges identify differences in amino-acid-sequence requirements for transactivation and ligand binding in the hormone-binding domain of the androgen receptor. These results provide new insights into ligand-dependent transactivation, and form a framework for the search for effective antagonists to be used in prostate-cancer therapy.

Amino Acid Sequence↗