epidermal-growth-factor and Prostatic-Hyperplasia

The data about the possible role of growth factors in pathogenesis of benign prostatic hyperplasia are presented in this review article. Special attention is paid to the basic fibroblast growth factor (bFGF) and transforming growth factor-beta 1 (TGF-beta1). The data about the regional concentration of growth factors in normal and benign hyperplastic human prostates are provided.

Topics: Aged; Epidermal Growth Factor; Fibroblast Growth Factors; Humans; Immunohistochemistry; Male; Muscle, Smooth; Prostatic Hyperplasia

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.

Topics: Animals; Cell Division; Disease Models, Animal; Epidermal Growth Factor; Fibroblast Growth Factors; Growth Substances; Humans; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Rats; Somatomedins; Transforming Growth Factors

Topics: Animals; Epidermal Growth Factor; Fibroblast Growth Factors; Growth Substances; Humans; Kidney Neoplasms; Male; Prostatic Hyperplasia; Prostatic Neoplasms; Somatomedins; Transforming Growth Factor beta; Urinary Bladder Neoplasms; Urogenital Neoplasms

A comment is made on the role growth factors play on the regulation of prostate growth. These factors require the mediation of an specific membrane receptor to which they have to bind in order to exercise their action correctly. The objective of the present job is to carry out a comprehensive review of each and every growth factor involved in prostate growth: family of the epidermal growth factor, family of the beta-transforming growth factor, and family of the fibroblast growth factor. As a final conclusion, it should be mentioned that the two prostate growth-regulating factors more extensively studied and with greater etiopathogenic relevance in benign prostate hyperplasia, are the epidermal growth factor and, more particularly, the fibroblast growth factor.

Topics: Epidermal Growth Factor; Fibroblast Growth Factors; Humans; Male; Prostatic Hyperplasia; Transforming Growth Factor beta

Topics: Animals; Cell Division; Dogs; Epidermal Growth Factor; Growth Substances; Humans; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Transforming Growth Factor beta

Growth and development of the prostate depend on androgen action and other factors. Androgens act directly by specific nuclear receptors and induce or control many effects mediated by peptide growth factors on both epithelial and stromal cells. The major important peptide growth factors detected in the prostate are Fibroblast Growth Factors, Transforming Growth Factors, and Epidermal Growth Factor. These factors are not prostate specific. They are produced by epithelial or stromal cells and regulate the growth of these two cell compartments through autocrine or paracrine pathways. Overproduction of these factors or modification in affinity or number of cell receptors may participate in the two main prostatic pathologies: benign hyperplasia or adenocarcinoma. Recently, other factors have been evidenced in the prostate: Interleukine 6, Nerve Growth Factor or Insulin-like Growth factors. The study of the localization and the effects of these factors may be crucial to understand the prostatic development and diseases and may suggest new targets for therapy.

Topics: Adenocarcinoma; Epidermal Growth Factor; Fibroblast Growth Factors; Humans; Interleukin-6; Male; Nerve Growth Factors; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Suramin; Transforming Growth Factor alpha; Transforming Growth Factor beta

Much research has been conducted to determine which tissue (epithelium or stroma) in the prostate gives rise to benign prostatic hyperplasia (BPH). Considering that BPH displays two structural compartments, stromal and epithelial and that the periurethral and transitional regions are particularly involved, the immunohistochemical and regional evaluation of steroid receptors concentration, 5 alpha reductase, DHT and estrogen activity, may show important data on the role of these factors in BPH development. We started a immunohistochemical study on the epidermal growth factor (EGF) concentrations in the periurethral, central and pericapsular zones of BPH samples, considering the stroma-epithelium ratio; investigations are performed on BPH patients submitted to transvesical prostatectomy. Considering that the periurethral zone is particularly involved in BPH, the presence of high concentration of growth factors in this region, may support the concept of their involvement in BPH.

Topics: Adult; Aged; Aged, 80 and over; Animals; Biomarkers; Connective Tissue; Dihydrotestosterone; Epidermal Growth Factor; Epithelium; Fibroblast Growth Factors; Humans; Male; Middle Aged; Prevalence; Prostate; Prostatic Hyperplasia; Receptors, Androgen; Rodentia; Transforming Growth Factor beta

Topics: Cell Division; Epidermal Growth Factor; Fibroblast Growth Factors; Growth Substances; Humans; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Transforming Growth Factor beta

Topics: 5-alpha Reductase Inhibitors; Androgens; Androstenes; Azasteroids; Carcinoma; Cell Division; Cells, Cultured; Dihydrotestosterone; Epidermal Growth Factor; Finasteride; Gonadotropin-Releasing Hormone; Growth Substances; Humans; Insulin-Like Growth Factor I; Male; Peptides; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Tumor Cells, Cultured

To analyse the expression of the epidermal growth factor (EGF) system in prostate tissue and secretions obtained from patients with benign prostatic hyperplasia (BPH) treated with or without finasteride (which primarily targets the androgen-sensitive secretory epithelial cells in the prostate, with little effect on basal epithelial and stromal cells).. The expression of the EGF system was evaluated by enzyme-linked immunosorbent assay and immunohistochemistry in samples of prostate tissue and secretions from patients with BPH randomized for treatment with finasteride or placebo for 3 months before surgery.. Prostate tissue expressed the EGF receptor (HER1) and HER2, and the ligands EGF, transforming growth factor alpha (TGFalpha), heparin-binding (HB) EGF, betacellulin and amphiregulin. Treatment with finasteride produced greater concentrations of amphiregulin (P < 0.05) than did placebo, did not change the level of TGFalpha, HER1 and HER2, and tended to decrease the concentration of EGF, betacellulin and HB-EGF in prostate tissue. Using immunohistochemistry, HER1 and TGFalpha were both localized to the basal epithelial cells, and there was a strong positive correlation among the tissue concentrations of HER1, HER2 and TGFalpha. Amphiregulin localized to the luminal secretory epithelium. Prostate secretions contained only EGF, which was at levels approximately 150 times higher than in prostate tissue; treatment with finasteride did not affect the concentration of EGF in prostate secretion.. There were only minor changes in the expression of TGFalpha, HER1 and HER2 after finasteride treatment. This may represent an important system for the continuous growth and homeostasis of the androgen-independent basal epithelial cells in the prostate.

Topics: Amphiregulin; Betacellulin; EGF Family of Proteins; Epidermal Growth Factor; ErbB Receptors; Finasteride; Glycoproteins; Growth Substances; Humans; Immunohistochemistry; Intercellular Signaling Peptides and Proteins; Male; Prospective Studies; Prostatic Hyperplasia; Receptor, ErbB-2; Transforming Growth Factor alpha

Benign prostatic hyperplasia (BPH) poses a significant health concern amongst elderly males. Canagliflozin (Cana), a selective sodium-glucose co-transporter 2 (SGLT2) inhibitor, has a powerful anti-inflammatory influence. Nevertheless, its role in treating BPH has not been clarified. Therefore, the study aimed to investigate the potential ameliorative effect of Cana on experimentally induced BPH in rats and explore the underlying mechanisms compared to the standard finasteride (Fin). The study employed histological analysis, biochemical assays using ELISA, and western blotting. Animals were categorized into four groups: Control (2.5 ml/kg CMC, orally + 3 ml/kg olive oil, subcutaneous), BPH (3 mg/kg testosterone, subcutaneous + CMC orally), Fin-treated BPH (5 mg/kg, orally), and Cana-treated BPH (5 mg/kg, orally), for 28 days. The BPH group showed obvious BPH manifestations including an increase in prostate weight (PW), prostate index (PI), dihydrotestosterone (DHT) level, and histological aberrations compared to control. Fin and Cana therapy had a comparable impact. Cana treatment significantly reduced PW and PI, besides it improved prostatic biochemical, and histopathological features compared to BPH, consistent with in silico study findings. Cana was associated with downregulation of the androgen axis, increased miR-128b expression, with a lowered expression of epidermal growth factor (EGF) and its receptor. Phosphorylation of STAT3 and its downstream proliferative markers were significantly reduced suggesting apoptotic activity. Cana markedly rescued the BPH-induced upregulation of IL-1β, and iNOS levels. Altogether, the current study demonstrates that Cana could impede BPH progression, possibly by modulating miR-128b/EGFR/EGF and JAK2/STAT3 pathways and downregulating AR, cyclin D1, and PCNA immunoreactivity.

Topics: Aged; Animals; Canagliflozin; Epidermal Growth Factor; ErbB Receptors; Finasteride; Humans; Hyperplasia; Janus Kinase 2; Male; MicroRNAs; Prostate; Prostatic Hyperplasia; Rats; Sodium-Glucose Transporter 2 Inhibitors; STAT3 Transcription Factor

In this study,mouse models of benign prostatic hyperplasia induced by subcutaneous injection of testosterone propionate was used to investigate the therapeutic effect and mechanism of Urtica hyperborean( UW) extracts on prostate hyperplasia in mice. The effects of UW extracts on prostate index,serum epidermal growth factor( EGF) and dihydrotestosterone( DHT) in model mice were observed,and the EGF and anti-apoptotic factor( Bcl-2) mRNA expression levels were detected as well as pathological changes in prostate tissue. The results showed that the ethyl acetate extraction and alcohol soluble fraction of the UW could significantly reduce the prostate index,reduce the serum DHT and EGF levels( P<0. 01),and significantly decrease the EGF and Bcl-2 mRNA expression( P<0. 01),significantly improved the morphological structure of prostate tissue. The above results confirmed that ethyl acetate extract and alcohol-soluble parts of UW have a good preventive effect on mice prostatic hyperplasia model,and its mechanism may be to reduce androgen levels by regulating polypeptide growth factors and/or inhibiting cell hyperproliferation and promoting apoptosis. This study laid the foundation for the further research on UW.

Topics: Animals; Dihydrotestosterone; Epidermal Growth Factor; Male; Medicine, Tibetan Traditional; Mice; Plant Extracts; Prostatic Hyperplasia; Proto-Oncogene Proteins c-bcl-2; Testosterone Propionate; Urticaceae

Stromal growth is critical for prostate enlargement during benign prostatic hyperplasia (BPH). While responses of prostate cells to single growth factors have been well characterized, responses to multiple growth factors at once are poorly understood. Here, we examined the effects of combinations between epidermal growth factor (EGF), fibroblast growth factor (FGF), and transforming growth factor-β1 (TGF-β1) in human prostate stromal cells.. EGF, FGF, and TGF-β1 were applied to WPMY-1 cells, an immortalized, non-malignant line of stromal cells from the human prostate. Hypertrophic responses were assessed by protein/DNA ratio, and cyclin D1 mRNA by RT-PCR. Expression of EGF, FGF, and TGF-β1 and their receptors in human prostate tissue was analyzed by RT-PCR, Western blot, and fluorescence staining.. Hypertrophic responses to single growth factors and combinations were similar. Combinations showed additive effects on cyclin D1 mRNA. Combination of EGF with TGF-β1, but not EGF or TGF-β1 alone, caused assembly of cells to a new two-dimensional structure, being characterized by dense aggregates connected by branches of few cells. EGF and TGF-β1 were detected together in human prostates. Receptors for EGF and TGF-β colocalized on stromal cells in human prostates.. Responses of prostate stromal cells to combinations of EGF, FGF, and TGF-β1 may be quantitatively different, qualitatively different, or similar to responses to single growth factors. The combination of EGF and TGF-β1, but not EGF or TGF-β1 alone, induces aggregation of prostate stromal cells, which may be relevant for morphogenesis.

Topics: Blotting, Western; Cell Line; Epidermal Growth Factor; Fibroblast Growth Factors; Fluorescence; Humans; Indoles; Male; Prostate; Prostatic Hyperplasia; Real-Time Polymerase Chain Reaction; Stromal Cells; Transforming Growth Factor beta1

This study was conducted to characterise the flavonoid components of total flavan glycoside from Abacopteris penangiana rhizomes (TFA) and its acid hydrolysate (AHT) through HPLC-DAD-ESI-MS/MS analysis, and to investigate the hypothesis that TFA and AHT exhibit anti-benign prostatic hyperplasia (BPH) potential in castrated rats with testosterone-induced BPH. HPLC-MS/MS analysis indicated that TFA is rich in flavan-4-ol glycosides and AHT mainly contains 3-deoxygenated anthocyanidin. After 4 weeks of administration, TFA and AHT successfully decreased the prostate index and prostate specific antigen plasma concentrations in the rats. Histoarchitectural improvement in the prostate gland was also observed. Reduced dihydrotestosterone, VEGF, bFGF, EGF, and KGF levels were observed both in TFA- and AHT-treated rats. Furthermore, the prostatic expression of Blc-2 was inhibited, whereas that of Bax and p53 was activated by TFA and AHT. In conclusion, TFA and AHT have anti-BPH properties. Hence, plants with flavan glycosides have potential use in the treatment of BPH.

Topics: Animals; Chromatography, High Pressure Liquid; Epidermal Growth Factor; Ferns; Flavonoids; Glycosides; Humans; Male; Mass Spectrometry; Plant Extracts; Prostatic Hyperplasia; Rats; Rats, Sprague-Dawley; Rhizome; Vascular Endothelial Growth Factor A

Inflammation plays a key role in the development of benign prostatic hyperplasia (BPH). Eicosanoids derived from the COX and 5-lipoxygenase (5-LOX) pathways are elevated in the enlarging prostate. Flavocoxid is a novel flavonoid-based 'dual inhibitor' of the COX and 5-LOX enzymes. This study evaluated the effects of flavocoxid in experimental BPH.. Rats were treated daily with testosterone propionate (3 mg·kg(-1)  s.c.) or its vehicle for 14 days to induce BPH. Animals receiving testosterone were randomized to receive vehicle (1 mL·kg(-1) , i.p.) or flavocoxid (20 mg·kg(-1) , i.p.) for 14 days. Histological changes, eicosanoid content and mRNA and protein levels for apoptosis-related proteins and growth factors were assayed in prostate tissue. The effects of flavocoxid were also tested on human prostate carcinoma PC3 cells.. Flavocoxid reduced prostate weight and hyperplasia, blunted inducible expression of COX-2 and 5-LOX as well as the increased production of PGE(2) and leukotriene B(4) (LTB(4) ), enhanced pro-apoptotic Bax and caspase-9 and decreased the anti-apoptotic Bcl-2 mRNA. Flavocoxid also reduced EGF and VEGF expression. In PC3 cells, flavocoxid stimulated apoptosis and inhibited growth factor expression. Flavocoxid-mediated induction of apoptosis was inhibited by the pan-caspase inhibitor, Z-VAD-FMK, in PC3 cells, suggesting an essential role of caspases in flavocoxid-mediated apoptosis during prostatic growth.. Our results show that a 'dual inhibitor' of the COX and 5-LOX enzymes, such as flavocoxid, might represent a rational approach to reduce BPH through modulation of eicosanoid production and a caspase-induced apoptotic mechanism.

Topics: Animals; Apoptosis; Arachidonate 5-Lipoxygenase; bcl-2-Associated X Protein; Caspase 9; Catechin; Cell Line, Tumor; Cell Survival; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dinoprostone; Drug Combinations; Epidermal Growth Factor; Humans; Leukotriene B4; Lipoxygenase Inhibitors; Male; Prostatic Hyperplasia; Rats; Rats, Sprague-Dawley; RNA, Messenger; Vascular Endothelial Growth Factor A

Peptide growth factors play an important role in several intracellular processes, such as cellular growth and differentiation, angiogenesis and apoptosis, as well as in carcinogenesis, since they contribute significantly to the malignant transformation. The prostate gland is abundant in growth factors. The two most known prostatic diseases, prostate cancer (PCa) and benign prostatic hyperplasia (BPH), are among the most common diseases that affect elderly men. This study was conducted using a quantitative real-time RT-PCR method in order to determine mRNA expression levels of peptide growth factors VEGF, FGF2, TGFB1, EGF, and IGF1 in tissue specimens from 42 patients with PCa, 42 with BPH, and 10 normal prostate samples obtained post-mortem from young individuals, in order to examine their association with prostatic hyperplasia and neoplasia. Our results show that in PCa, growth factors VEGF, EGF and FGF2 are overexpressed, while TGFB1 and IGF1 have reduced mRNA levels. In BPH, transcript levels of FGF2 and EGF are normal, while VEGF, TGFB1 and IGF1 exhibit downregulation. Further statistical analysis revealed that PCa patients with high levels of PSA blood levels have decreased FGF2 expression (p=0.016). Additionally, cancer patients with low Gleason score (<7) have increased EGF (p=0.035) and IGF1 (p=0.031) mRNA levels. IGF1 levels are also elevated in tumors with TNM stages T1-T2 (p=0.030). In BPH, older patients have reduced EGF expression (p=0.018), while IGF1 is overexpressed in younger patients (p=0.041). Additionally, the co-expression pattern of the five studied growth factors differs significantly among normal, benign and malignant prostate. These results implicate VEGF, FGF2, TGFB1, EGF and IGF1 in the development of both PCa and BPH, rendering them potential targets for disease detection, monitoring and therapy.

Topics: Adult; Aged; Aged, 80 and over; Epidermal Growth Factor; Fibroblast Growth Factor 2; Gene Expression Regulation, Neoplastic; Humans; Insulin-Like Growth Factor I; Male; Middle Aged; Prostatic Hyperplasia; Prostatic Neoplasms; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

To investigate the effects of epidermal growth factor (EGF) on estrogen receptor (ER) and androgen receptor (AR) in mouse prostate cells and explore the putative role of EGF in prostatic hyperplasia.. Sixty male Kunming mice were randomly divided into two EGF groups and one control group (n=20) and subjected to subcutaneous injection of 1 and 2 microg/day EGF and distilled water, respectively, for 28 consecutive days. The cellular expression of ER and AR in the prostate of mice in different groups was evaluated by flow cytometry.. Compared with the control group, the positivity rate of ER and its expression level were significantly increased in the mouse prostate after EGF treatment (P<0.01), and the ER expression level was significantly higher in mouse with 2 microg/day EGF treatment than in those treated with 2 microg/day EGF (P<0.01). AR positivity rate and expression level also increased significantly in comparison with the control group (P<0.05), but no significant variation was found between 1 microg/day and 2 microg/day EGF groups.. EGF can increase the cellular expression of ER and AR in mice prostate and may play a role in the pathogenesis of prostatic hyperplasia.

Topics: Animals; Epidermal Growth Factor; Flow Cytometry; Injections, Subcutaneous; Male; Mice; Prostate; Prostatic Hyperplasia; Random Allocation; Receptors, Androgen; Receptors, Estrogen

The human prostate gland is a well known source of H1 and H2 relaxin but information is lacking on the expression and potential role of the INSL3 peptide hormone within the prostate gland. In the present study we have investigated the expression of human INSL3 in patients with benign prostate hyperplasia (BPH), prostate intraepithelial neoplasia (PIN) and prostate carcinoma tissues. Of the prostate epithelial cells, strongest INSL3 expression was detected in the basal epithelial cell compartment. Weaker INSL3 mRNA expression and immunoreactive INSL3 production were observed in secretory epithelial cells and in interstitial smooth muscle cells. Prostate epithelial cells were also a source for transcripts encoding the INSL3 receptor LGR8 suggesting the presence of an autocrine/paracrine INSL3-LGR8 ligand-receptor system within the human prostate. Three human prostate carcinoma cell lines displayed differential gene activity for INSL3 and LGR8. While LNCaP was devoid of INSL3, the androgen-insensitive PC-3 and the stromal prostate cell line hPCP co-expressed INSL3 and LGR8 transcripts. In addition to expressing INSL3 mRNA, the LGR8-negative DU-145 also expressed an INSL3 splice form formerly demonstrated in thyroid carcinoma cells. When incubated with recombinant INSL3, PC-3 cells showed significantly increased intracellular cAMP levels indicating functional LGR8 receptors. INSL3 did not alter the proliferative or metabolic activity of PC-3 carcinoma cells. Instead, PC-3 responded to INSL3 with significantly enhanced tumor cell motility and a transcriptional down-regulation of ErbB receptors and EGF. All-trans-retinoic acid was demonstrated in PC-3 to up-regulate LGR8 gene activity in a dose- and time-dependent manner while having no effect on INSL3 gene activity. In conclusion, we have identified a functional INSL3-LGR8 ligand-receptor system in human prostate carcinoma cells.

Topics: Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cyclic AMP; Dose-Response Relationship, Drug; Epidermal Growth Factor; Epithelial Cells; Gene Expression; Humans; Immunohistochemistry; In Situ Hybridization; Insulin; Male; Prostatic Hyperplasia; Prostatic Intraepithelial Neoplasia; Prostatic Neoplasms; Proteins; Receptors, G-Protein-Coupled; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Time Factors; Tretinoin

The disintegrin metalloprotease ADAM-10 is a multidomain metalloprotease that is potentially significant in tumor progression due to its extracellular matrix-degrading properties. Previously, ADAM-10 mRNA was detected in prostate cancer (PCa) cell lines; however, the presence of ADAM-10 protein and its cellular localization, regulation, and role have yet to be described. We hypothesized that ADAM-10 mRNA and protein may be regulated by growth factors such as 5alpha-dihydrotestosterone, insulin-like growth factor I, and epidermal growth factor, known modulators of PCa cell growth and invasion.. ADAM-10 expression was analyzed by in situ hybridization and immunohistochemistry in prostate tissues obtained from 23 patients with prostate disease. ADAM-10 regulation was assessed using quantitative reverse transcription-PCR and Western blot analysis in the PCa cell line LNCaP.. ADAM-10 expression was localized to the secretory cells of prostate glands, with additional basal cell expression in benign glands. ADAM-10 protein was predominantly membrane bound in benign glands but showed marked nuclear localization in cancer glands. By Western blot, the 100-kDa proform and the 60-kDa active form of ADAM-10 were synergistically up-regulated in LNCaP cells treated with insulin-like growth factor I plus 5alpha-dihydrotestosterone. Epidermal growth factor also up-regulated both ADAM-10 mRNA and protein.. This study describes for the first time the expression, regulation, and cellular localization of ADAM-10 protein in PCa. The regulation and membrane localization of ADAM-10 support our hypothesis that ADAM-10 has a role in extracellular matrix maintenance and cell invasion, although the potential role of nuclear ADAM-10 is not yet known.

Topics: ADAM Proteins; ADAM10 Protein; Amyloid Precursor Protein Secretases; Androgens; Blotting, Western; Cell Membrane; Cell Nucleus; Dihydrotestosterone; Epidermal Growth Factor; Gene Expression Regulation; Humans; Immunoenzyme Techniques; In Situ Hybridization; Insulin-Like Growth Factor I; Male; Membrane Proteins; Metalloendopeptidases; Prostatic Hyperplasia; Prostatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Tumor Cells, Cultured

Benign prostatic hyperplasia (BPH) is a condition that affects a significant population of older males, yet its pathogenesis is not clearly understood. This study was designed to give broader insight into the early development of BPH by looking at changes in growth factor production in the ventral prostate. To accomplish this, Sprague Dawley rats (n = 16, 250-300 g) were randomly divided into four equal groups. Three treatment groups were each implanted with ceramic drug delivery devices that were designed to deliver continuous physiologic doses of testosterone (T), dihydrotestosterone (DHT), and androstenedione (AED) for ninety-day duration. Immuno-histochemical analysis for epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) indicated whether these growth factors were involved in early processes of BPH induced by the delivery of androgens. Histopathological evaluation demonstrated increased positive reactivity for both EGF and bFGF in all steroid treated animals compared to controls. A similar trend was observed in the vascular endothelium. This information could be helpful in developing new methods for early diagnosis of BPH, but more importantly this knowledge provides the literature with clues about the cellular responses encountered at the initiation of the disease process.

Topics: Androgens; Androstenedione; Animals; Calcium Phosphates; Dihydrotestosterone; Drug Implants; Endothelium, Vascular; Epidermal Growth Factor; Fibroblast Growth Factors; Male; Prostate; Prostatic Hyperplasia; Rats; Reference Values; Testosterone

Prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA) are the markers of human prostatic gland. However, it is still not completely understood if and how, steroid hormones and growth factors affect their expression and metabolism in the respect to the major pathologies of the gland. Appropriate studies were carried out on histopathologically diagnosed benign prostatic hyperplasia--BPH (n = 42) using tissue slices and cells derived from them. They were incubated with steroid hormones: 5-alpha-dihydrotestosterone (DHT), estradiol (E) and growth factors: epidermal growth factor (EGF), basic fibroblastic growth factor (bFGF) under culture conditions for up to 24 hours. P-labelled specific oligonucleotide probes were used to analyze total RNA isolated from each sample for the presence of PAP and PSA mRNAs. DHT, E, bFGF, EGF or both DHT + bFGF and DHT + EGF increased PAP and PSA mRNA levels in a time- and dose-dependent manner. The highest and statistically significant increase (P < 0.001) for PAP mRNA was observed when DHT + bFGF were present in the medium while for PSA mRNA if DHT + EGF were added to the medium. Slow but constant decrease of PAP and PSA mRNA levels was observed in the absence of each of these factors in the incubation medium. The results suggest that early expression of PSA and PAP genes and/or their mRNA stability strongly depend on DHT while differ in their response to EGF and bFGF.

Topics: Acid Phosphatase; Base Sequence; Blotting, Northern; Dihydrotestosterone; Epidermal Growth Factor; Estradiol; Fibroblast Growth Factor 2; Growth Substances; Humans; Male; Molecular Sequence Data; Prostate; Prostate-Specific Antigen; Prostatic Hyperplasia; Protein Tyrosine Phosphatases; RNA, Messenger; Steroids; Tumor Cells, Cultured

The physiological mechanisms by which soluble mediators of cell proliferation and survival alter expansion of the prostatic stroma in benign prostatic hyperplasia are poorly understood. We recently identified heparin-binding epidermal growth factor like growth factor (HB-EGF) as a product predominantly of the smooth muscle cell compartments of the adult human prostate. We assess the potential role of this growth factor as a stromal cell regulator.. Primary cultures of desmin and alpha-actin positive human prostate stromal cells were shown to express several cell associated HB-EGF isoforms as well as the primary cognate HB-EGF receptor, ErbB1/HER1, suggesting the existence of an autocrine or juxtacrine regulatory loop. The related receptor tyrosine kinase, ErbB2/HER2, was also expressed as assessed by reverse transcriptase (RT) polymerase chain reaction (PCR). HB-EGF messenger RNA levels in human prostate stromal cells increased modestly (70%) in response to a repetitive mechanical stimulus, a lower response than has been reported for neonatal rat bladder smooth muscle cells, in which HB-EGF was originally identified as a mechanically responsive gene.. HB-EGF, epidermal growth factor and basic fibroblast growth factor stimulated human prostate stromal cell growth, while a specific antagonist of HB-EGF, [Glu52]-diphtheria toxin/CRM197, inhibited human prostate stromal growth in serum-free medium by a mechanism that did not involve increased apoptosis. A function blocking antibody against CD9/DRAP27/MRP-1, a cell surface binding partner of the membrane form of HB-EGF, also stimulated human prostate stromal cell proliferation.. HB-EGF is an endogenously produced human prostate stromal cell growth factor and, thus, may have a role as a physiologically relevant autocrine or juxtacrine mediator of stromal expansion in benign prostatic hyperplasia.

Topics: Adult; Animals; Animals, Newborn; Cell Division; Epidermal Growth Factor; Fibroblast Growth Factor 2; Heparin; Heparin-binding EGF-like Growth Factor; Humans; In Vitro Techniques; Intercellular Signaling Peptides and Proteins; Male; Prostate; Prostatic Hyperplasia; Rats; Stromal Cells

To study the molecular mechanism of rat prostate atrophy induced by epristeride.. MTT test was used to determine the effect of epristeride on the growth of prostatic epithelial cell induced by exogenous epithelial growth factor (EGF) or insulin-like growth factor-I (IGF-I). RT-PCR and flow cytometry were then used to quantitatively detect the mRNA and protein expressions of EGFR and IGF-I R of the epithelial cells treated or untreated with epristeride.. Epristeride attenuated growth of epithelial cells induced by exogenous EGF, IGF-I. Epristeride 360 nmol/L inhibited EGFR and IGF-I R expression at mRNA level, while epristeride 180 nmol/L had no marked effect on EGFR and IGF-I R mRNA expression. Both epristeride 180 nmol/L and 360 nmol/L could down regulate EGFR and IGF-I R protein levels.. The molecular mechanisms of prostatic epithelial cell atrophy induced by epristeride might be associated with alteration in the expression of growth factor receptors such as EGF and IGF-I.

Topics: 5-alpha Reductase Inhibitors; Androstadienes; Animals; Cell Division; Cells, Cultured; Epidermal Growth Factor; Epithelial Cells; ErbB Receptors; Insulin-Like Growth Factor I; Male; Prostate; Prostatic Hyperplasia; Rats; Receptor, IGF Type 1; RNA, Messenger

The Epidermal Growth Factor (EGF) plays an important role in the regulation of in vitro growth of prostate cells inducing a strong mitogenic effect. Nevertheless in our previous study we observed that the treatment of human hypertrophic prostate cell line U285 with exogenous EGF produces a restricted effect on the cellular growth rate. This phenomenon could be due to the capacity of the cells to produce EGF. In this study we aimed to verify this hypothesis by evaluating the presence of mRNA of EGF and EGF receptor (EGF-R) and of their translation products in U285 cells, before and after the treatment with suramin and exogenous EGF. Moreover we studied the effects exerted by these substances on the proliferative rate of the cells U285 after different treatment protocols. The presence in the cells of mRNA for EGF and EGF-R and of their translation products was demonstrated by means of reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods respectively. The modification of growth rate induced by these drugs was studied by FRAME Cytotoxicity Test. The operative modalities adopted to carry out these growth assays tended to 1) focus the effects of suramin in relation to in vitro cellular growth phase; 2) verify the reversibility of its effects; 3) ascertain if it was possible to antagonize the action of suramin by adding exogenous EGF. The results obtained from the RT-PCR showed the presence, in the control cells and in the treated ones, of mRNA coding for EGF and EGF-R. The immunocytochemical analysis indicated that 20% of the control cells are EGF positive, and 83% are EGF-R positive, confirming the results obtained with RT-PCR. Moreover, these stainings showed that the treatment with EGF does not significantly modify the percentage of cells marked by the anti-EGF antibody, while treatments with suramin and suramin plus EGF double this percentage. None of the treatments modifies the percentage of EGF-R positive cells. The growth assays showed that the exposition to highest doses of suramin in the first 24 h of cultures causes a decrease (p < 0.05) of the cellular proliferation during the following 48 h and 72 h and that these effects are irreversible. Moreover, a contemporaneous exposition of the cells to EGF and suramin at seeding strengthens the cytotoxic action of the last drug. To sum up, the demonstration of the presence in the U285 cells of mRNA coding for EGF and EGF-R and of the corresponding proteins, confirms the hyp

Topics: Analysis of Variance; Cell Division; Cell Line; Cell Survival; Epidermal Growth Factor; ErbB Receptors; Humans; Immunohistochemistry; Male; Prostate; Prostatic Hyperplasia; Protein Biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Suramin

Except for prostate volume, little is known about the factors influencing serum prostate specific antigen (PSA) levels. Considering that dihydrotestosterone and epidermal growth factor are regulators of the proliferation and differentiation in the epithelial component of human prostate tissue and that PSA is produced only by the epithelial cells of the gland, studies were performed on patients with a histological diagnosis of benign prostatic hyperplasia (BPH) to establish whether a significant association exists between the intraprostatic concentration of dihydrotestosterone or epidermal growth factor and serum PSA levels.. A total of 20 patients with BPH who had not been previously treated were part of a larger study on the correlation among PSA, prostate volume and age, and were evaluated according to the algorithm in the guidelines of the international consultation on BPH. All men underwent open suprapubic prostatectomy to enucleate the entire adenoma and in each case sections were made in the periurethral, subcapsular and intermediate zones of the BPH tissue. Dihydrotestosterone and epidermal growth factor concentrations were evaluated by radioimmunoassay in the periurethral zone and in total BPH tissue.. In these 20 patients with BPH serum PSA levels were significantly associated with epidermal growth factor but not with dihydrotestosterone concentrations in total BPH tissue (r = 0.7762, p = 0.00002836 and r = 0.3923, p = 0.0956307, respectively). A stronger association was found between PSA levels and the periurethral concentration of epidermal growth factor and dihydrotestosterone (r = 0.8117, p = 0.000005 and r = 0.5656, p = 0.0098326, respectively). On the contrary, epidermal growth factor and dihydrotestosterone were not significantly associated with prostate volume (p = 0.957415 and p = 0.531439, respectively).. To our knowledge this study is the first report in the literature to demonstrate an association between serum PSA, and dihydrotestosterone and epidermal growth factor levels, particularly in the periurethral zone of human BPH tissue. These data suggest the importance of epidermal growth factor and dihydrotestosterone in influencing serum PSA levels.

Topics: Aged; Dihydrotestosterone; Epidermal Growth Factor; Humans; Linear Models; Male; Middle Aged; Prostate; Prostate-Specific Antigen; Prostatic Hyperplasia

Although growth factors such as epidermal growth factor (EGF), transforming growth factor (TGF)-alpha, and TGF-beta are important regulators of prostate cell growth in vitro and in animal models, evidence to support their role in human prostate cancer development remains sparse. We previously showed that men without prostate cancer have concentrations of EGF and TGF-alpha in expressed prostatic fluid (EPF) that are individually distinct and stable over time. This study addressed whether growth factor levels in EPF are associated with the presence or progression of prostate cancer.. We measured levels of immunoreactive EGF, TGF-alpha, and TGF-beta1 in stored EPF samples from three age-matched groups: 19 men with untreated, histologically diagnosed prostate cancer (CaP), 38 with benign prostate hyperplasia (BPH), and 19 with normal prostate glands (NPD).. Median TGF-alpha was lower in the BPH group (0.45 ng/ml) than in either CaP (0.63 ng/ml) or NPD (0.58 ng/ml) groups (P = 0.03 and 0.12, respectively). For EGF, the median was lowest in the CaP group and highest in the NPD group (92.5 ng/ml vs. 175.5 ng/ml, P = 0.006). For TGF-beta1, the median level in CaP was 2.7 times higher than the median level among all controls (6.65 ng/ml vs. 2.46 ng/ml, P = 0.002). Growth factor levels were not associated with tumor stage or Gleason score. However, the single case with distant metastases had TGF-beta1 levels 23-fold higher than the CaP median.. The results suggest that at the time of CaP diagnosis, EGF levels in EPF are significantly lower, and TGF-beta1 levels significantly higher, than normal. Marked overexpression of TGF-beta1 in advanced CaP might be reflected in extremely high EPF levels.

Topics: Aged; Disease Progression; Epidermal Growth Factor; Growth Substances; Humans; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Radioimmunoassay; Transforming Growth Factor alpha; Transforming Growth Factor beta

Immunoreaction to TGF-alpha was limited to the basal epithelial cells of focal areas in the normal prostates. In benign prostatic hyperplasia (BPH) the immunostained areas were more widespread and immunolabelling was observed in both basal and columnar (secretory) cells of the epithelium. Some cells in the connective tissue stroma were also stained. In prostatic adenocarcinoma, epithelial immunostaining was even more extensive and intense than in BPH, and some stromal cells were also stained. Epidermal growth factor (EGF) immunostaining was only present in some basal cells in normal prostates. In BPH, this immunoreaction was strong in the basal cells and even stronger in the secretory cells. In prostatic cancer, the intensity of epithelial cell immunoreactivity was intermediate between that of normal prostates and that of BPH specimens. EGF-receptor immunostaining was focal and located in the basal cells in normal prostates. In BPH, labelling was also localized in basal cells but extended to wider areas. Some stromal cells appeared weakly labelled. In the prostatic carcinoma, both basal and columnar cells appeared stained and the number of immunolabelled stromal cells was higher than in BPH. The results presented suggest that, in normal conditions, EGF and TGF-alpha act as autocrine growth factors for the basal cells of the prostatic epithelium. In BPH this action is maintained and, in addition, the columnar cells start to secrete both factors which are bound by the basal cell receptors, giving rise to a paracrine regulation which probably overstimulates basal cell proliferation. In prostatic carcinoma, besides these regulatory mechanisms, the acquisition of EGF-receptors by the secretory cells develops an autocrine regulation which might induce their proliferation.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Animals; Antibody Specificity; Connective Tissue; Epidermal Growth Factor; Epithelial Cells; ErbB Receptors; Humans; Immunoenzyme Techniques; Male; Mice; Middle Aged; Neoplasm Proteins; Organ Specificity; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Rabbits; Stromal Cells; Transforming Growth Factor alpha

To assess the effect of the lipidosterolic extract of Serenoa repens (LSESr) on in vitro cell proliferation in biopsies of human prostate. Cell proliferation was assessed by incorporation of [3H]thymidine followed by historadiography.. Basic fibroblast growth factor (b-FGF) induced a considerable increase in human prostate cell proliferation (from +100 to +250%); the glandular epithelium was mainly affected, minimal labeling being recorded in the other regions of the prostate. Similar results were observed with epidermal growth factor (EGF), although the increase in cell proliferation was not recorded in some cases. Lovastatin, an inhibitor of hydroxymethylglutaryl coenzyme A, antagonized both the basal proliferation and the growth factor-stimulated proliferation of human prostate epithelium (EGF, mean inhibition approximately 80-95%; b-FGF, mean inhibition approximately 40-90%). Geraniol, a precursor of both farnesyl pyrophosphate and geranylgeranyl pyrophosphate, and farnesol, the precursor of farnesyl pyrophosphate, increased cell proliferation only in some prostate specimens, this effect being antagonized by lovastatin. LSESr did not affect basal prostate cell proliferation, with the exception of two prostate specimens in which a significant inhibition of basal proliferation was observed with the highest concentration of LSESr (30 micrograms/ ml). In contrast, LSESr inhibited b-FGF-induced proliferation of human prostate cell cultures; this effect was significant for the highest concentration of LSESr (30 micrograms/ml). In some prostate samples, a similar inhibition was also noted with lower concentrations. Unsaturated fatty acids (UFA), in the range 1-30 ng/ml), did not affect the basal prostate cell proliferation, only a slight increase in cell proliferation was noted in 1 prostate specimen. UFA (1, 10 or 30 micrograms/ml) markedly inhibited the b-FGF-induced cell proliferation down to the basal value. Lupenone, hexacosanol and the unsaponified fraction of LSESr markedly inhibited the b-FGF-induced cell proliferation, whereas a minimal effect on basal cell proliferation was noted.. Despite the large variability in the response of the prostate samples to b-FGF, these results indicate that LSESr and its components affect the proliferative response of prostate cells to b-FGF more than their basal proliferation.

Topics: Acyclic Monoterpenes; Androgen Antagonists; Cell Division; Cells, Cultured; Epidermal Growth Factor; Farnesol; Fibroblast Growth Factor 2; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Lovastatin; Male; Plant Extracts; Prostate; Prostatic Hyperplasia; Serenoa; Terpenes

It is well recognized that the actions of androgens alone do not explain the hyperplastic development of the gland that occurs in elderly men. The increasing number of reports confirming the lack of mitogenic activity of androgens coupled with the powerful mitogenic activity of growth factors and the discovery of growth factor receptors led to an increased interest in the putative role of growth factors in prostate physiopathology. We have previously demonstrated the presence and the cellular localization of epidermal growth factor and of the related peptide, transforming growth factor-alpha, together with their common receptor in the epithelial compartment of the human hyperplastic prostate tissue (BPH). In the present study we examined the expression and cellular localization of messenger ribonucleic acid (RNA) encoding keratinocyte growth factor (KGF) and its receptor in human hyperplastic prostate tissue. RT-PCR of total RNA extracted from BPH tissues documented the presence of transcripts for KGF and its receptor. In situ hybridization with specific RNA probes synthesized from the respective complementary DNA demonstrated that KGF mRNA was mainly localized in the stromal cells, whereas its receptor was mainly localized in the prostate epithelium. Moreover, the mitogenic activity of KGF on cultured BPH cells compared to that of other growth factors has been tested. Our findings clearly indicate that KGF has the ability to function as a potent mitogen in BPH cells. Our data support the hypothesis that KGF plays an important role in prostate growth and that in human prostate it seems to act in a paracrine fashion.

Topics: Cell Division; Epidermal Growth Factor; Fibroblast Growth Factor 10; Fibroblast Growth Factor 2; Fibroblast Growth Factor 7; Fibroblast Growth Factors; Gene Expression; Growth Substances; Humans; In Situ Hybridization; Male; Polymerase Chain Reaction; Prostatic Hyperplasia; Receptor, Fibroblast Growth Factor, Type 2; Receptors, Fibroblast Growth Factor; Receptors, Growth Factor; RNA-Directed DNA Polymerase; RNA, Messenger

The n-hexane lipido-sterol extract of Serenoa repens (LSESr, Permixon, Pierre Fabre Medicament, Castres, France), a phytotherapeutic agent used in the treatment of benign prostatic hyperplasia (BPH), has a multisite mechanism of action including inhibition of types 1 and 2 5alpha-reductase and competitive binding to androgen receptors in prostatic cells. Here, the response of testosterone (T), dihydrotestosterone (DHT), and epidermal growth factor (EGF) in BPH tissue of patients treated with LSESr (320 mg/day for 3 months) is analyzed.. BPH samples were sectioned in periurethral, subcapsular, and intermediate regions: in each region T, DHT, and EGF were determined by radioimmunoassay after purification on celite columns or Sep-pak C18 cartridges.. In the untreated group, T, DHT, and EGF presented the highest concentrations in the periurethral region (615 +/- 62 (SE) pg/g tissue, 7,317 +/- 551 pg/g tissue, and 20.9 +/- 3.3 ng/g tissue, respectively) with respect to the peripheral subcapsular region (425 +/- 45 pg/g tissue, 4,215 +/- 561 pg/g tissue, and 10.8 +/- 1.4 ng/g tissue, respectively). In the LSESr-treated group, a statistically significant reduction was observed, mainly in the periurethral region of DHT (2,363 +/- 553 pg/g tissue, P < 0.001) and EGF (6.98 +/- 2.48 ng/g tissue, P < 0.01), with increased T values (1,023 +/- 101 pg/g tissue, P < 0.001).. The decrease of DHT and the rise of T in BPH tissue of patients treated with Permixon confirms the capacity of this drug to inhibit in vivo 5alpha-reductase in human pathological prostate. A marked decrease of EGF, associated with DHT reduction, was also observed. These biochemical effects, similar to those obtained with finasteride, are particularly evident in the periurethral region, whose enlargement is responsible for urinary obstruction, with respect to the subcapsular region. A possible speculation is that the preferential reduction of DHT and EGF content in the periurethral region is involved in the clinical improvement of the obstructive symptoms in BPH during LSESr therapy.

Topics: Aged; Androgen Antagonists; Cholestenone 5 alpha-Reductase; Dihydrotestosterone; Epidermal Growth Factor; Humans; Male; Middle Aged; Oxidoreductases; Plant Extracts; Prostatic Hyperplasia; Serenoa; Testosterone

A variety of hormones have demonstrated effects on prostatic tissue growth dynamics. Our goal was to define the effect of dihydrotestosterone (DHT), epidermal growth factor (EGF), and prolactin (PRL) on prostate cellular proliferation.. Thirty benign human prostatic hyperplasias (BPH) were maintained 48 hr as in vitro cultures. Culture media were supplemented with EGF, DHT, and PRL alone and in combinations. Proliferation was assessed by labeling with tritiated thymidine.. The proliferative response of individual BPH cultures was heterogeneous. DHT and EGF tended to have a greater proliferative effect than PRL, both in terms of the percent cultures responding and the magnitude of the response. PRL antagonized EGF-induced proliferative effects. EGF- and PRL-mediated effects correlated with each other, while DHT-mediated effects did not correlate with either those of PRL or EGF.. The proliferative response of individual BPH to DHT, EGF, and PRL, alone or in combination, is too variable to define a predictable response to their influence. Our methodology represents a technique with the capacity to define therapeutic potential for individual cases.

Topics: Aged; Aged, 80 and over; Cell Cycle; Cell Division; Cells, Cultured; Dihydrotestosterone; DNA; Drug Synergism; Epidermal Growth Factor; Humans; Male; Middle Aged; Prolactin; Prostate; Prostatic Hyperplasia; Thymidine; Time Factors

This work studies the effects of dihydrotestosterone (DHT) and epidermal growth factor (EGF) on the growth, morphology and phenotype characterisation of the U285 line obtained from human prostate hyperplastic tissue. Modifications of growth rate induced by these two substances have been evaluated by means of the neutral red assay formulated by Borenfreund and Puerner (1985) as well as by means of Kenacid blue assay described by Knox et al. (1986), culturing cells for 24, 48 and 72 hr with scalar doses of DHT (0.5, 1, 2, 5, 10 microM) and EGF (5, 10, 20, 100 ng/ml). An optical microscope connected to a computer aided system and a scanning electron microscope were used to monitor morphological changes induced by DHT and EGF. The immunophenotype characterisation of the treated and control cells was carried out by using a monoclonal antibody panel. Our results show that the expression of anti-cytokeratin 5+6+18, anti-cytokeratin 8+18+19 and anti-proline-4-hydroxylase antibodies varied in relation to the type of treatment undergone by the cells. Moreover, exogenous DHT does provoke a flattening of the U285 cells without modifying their rate of growth, while EGF both shortens the lag phase reactivating the quiescent cells and regulates the subsequent log growth phase, thus causing no cellular overgrowth.

Topics: Cell Division; Cells, Cultured; Dihydrotestosterone; Dose-Response Relationship, Drug; Epidermal Growth Factor; Humans; Immunophenotyping; Male; Microscopy, Electron, Scanning; Middle Aged; Prostatic Hyperplasia

Prostatic fluid (PF) provides a unique medium for noninvasive evaluation of critical growth and differentiation signals in the prostatic microenvironment. The purpose of this study was to establish the feasibility of measuring two prostatic mitogens, epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) in PF, and specifically to quantify extraneous variability attributable to the assay itself, sample handling, or biological variation within an individual over time.. PF was collected by transrectal massage from consecutive patients attending a urology clinic. Pooled PF and individual samples from 25 men with stable benign prostatic hyperplasia (BPH) were analyzed for EGF and TGF-alpha by radioimmunoassay and for total protein.. Reproducibility was adequate at dilutions as low as 1:50 (2-microliter pooled sample) and 1:5 (20 microliters) for EGF and TGF-alpha, respectively. Results were not affected by freeze-thaw cycles, time in storage, or protease inhibition in fresh PF. EGF and TGF-alpha were detectable in 100% and 92% of individual men, with respective means of 152 and 0.2 ng/ml. Correlations between two samples obtained from the same man within 12 months were highly significant (EGF r = 0.89, TGF-alpha r = 0.71). Protein concentrations were consistent over time; expression of either peptide per weight of protein rather than per volume did not improve within-man correlation. Between-man variability far exceeded within-man variability for both peptides, and was estimated to account for 84% and 61% of the total variability in EGF and TGF-alpha, respectively. There was no correlation between EGF and TGF-alpha in the same samples.. We conclude that men with BPH secrete consistent and distinct levels of EGF-related peptides in PF, and that these levels can be detected with acceptable sensitivity and precision by radioimmunoassay (RIA). Measurement of TGF-alpha, which has not been reported previously, requires a relatively larger sample.

Topics: Analysis of Variance; Body Fluids; Epidermal Growth Factor; Feasibility Studies; Humans; Male; Mitogens; Prostate; Prostatic Hyperplasia; Radioimmunoassay; Reproducibility of Results; Transforming Growth Factor alpha

The potent vasoconstrictor endothelin-1 (ET-1) is at its highest concentration in the normal human ejaculate and is associated with the progression of metastatic prostate cancer. ET-1 protein expression is detected in situ in 14 of 14 primary cancers and 14 of 16 metastatic sites of human prostatic carcinoma. Exogenous ET-1 induces prostate cancer proliferation directly and enhances the mitogenic effects of insulin-like growth factor I, insulin-like growth factor II, platelet-derived growth factor, basic fibroblast growth factor, and epidermal growth factor in serum-free conditions in vitro. The ETA-selective receptor antagonist A-127722 inhibits ET-1-stimulated growth, but the ETB-selective receptor antagonist BQ-788 does not. ET-3, an ETB-selective agonist, also had no effect on prostate cancer growth. No specific ETB-binding sites could be demonstrated in any established human prostate cancer cell line tested, and ETB mRNA, detected by reverse transcription PCR, was reduced. The predominance of ETB binding on human benign prostatic epithelial tissue is not present in metastatic prostate cancer by autoradiography. In human prostate cancer progression to metastases, ET-1 and ETA expression are retained, whereas ETB receptor expression is reduced.

Topics: Apoptosis; Atrasentan; Base Sequence; Cell Division; Cell Line; Culture Media, Serum-Free; DNA Primers; DNA, Neoplasm; Endothelin Receptor Antagonists; Endothelins; Epidermal Growth Factor; Fibroblast Growth Factor 2; Gene Expression; Growth Substances; Humans; Immunohistochemistry; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Male; Mitotic Index; Molecular Sequence Data; Neoplasm Metastasis; Platelet-Derived Growth Factor; Polymerase Chain Reaction; Prostate; Prostatectomy; Prostatic Hyperplasia; Prostatic Neoplasms; Pyrrolidines; Receptor, Endothelin B; Receptors, Endothelin; RNA, Messenger; Tumor Cells, Cultured

Parathyroid hormone-related protein (PTHrP) has previously been shown to be expressed in human prostatic tissue and in prostatic cancer cell lines. In the present study, PTHrP immunoreactivity was detected in the glandular epithelium of normal prostate and benign prostatic hyperplasia (BPH), as well as in prostatic adenocarcinoma (CaP). Epithelial cell cultures derived from normal, BPH, and CaP tissues were also stained by antibodies against PTHrP, and northern analysis revealed multiple transcripts of PTHrP in the cellular RNA. PTHrP (1-34) was measurable by radioimmunoassay (RIA) in media conditioned by the prostatic epithelial cell cultures, and PTHrP accumulated in conditioned media during a 72 hr time course. Addition of complete growth medium to starved cells resulted in increased PTHrP mRNA levels by 1 hr, with maximal stimulation at 8-24 hr. Several individual factors contained in the complete growth medium were tested for their ability to regulate PTHrP expression. Epidermal growth factor (EGF) was the major inducer of PTHrP expression, while cholera toxin, bovine pituitary extract, hydrocortisone, and insulin had minimal or no effect on PTHrP transcript levels. Since each of these factors is growth stimulatory, the unique ability of EGF to induce PTHrP is apparently unrelated to mitogenicity. 1,25-Dihydroxyvitamin D3[1,25(OH)2D3], an inhibitor of PTHrP expression in several other cell types, had no effect on steady-state levels of PTHrP mRNA expressed by epithelial cells in complete growth medium, although prostate cells have vitamin D receptors and are responsive to 1,25(OH)2D3 in other ways. Our results indicate that PTHrP expression is not confined to the neuroendocrine cells of the human prostate and that our culture system can be used as a model to investigate the role of PTHrP in the prostate.

Topics: Adenocarcinoma; Calcitriol; Cells, Cultured; Culture Media; Epidermal Growth Factor; Epithelium; Gene Expression Regulation; Humans; Immunohistochemistry; Kinetics; Male; Parathyroid Hormone-Related Protein; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Proteins; RNA, Messenger

The expression of epidermal growth factor receptor (EGF-R), transforming growth factor alpha (TGFalpha), and epidermal growth factor (EGF) was evaluated in a series of prostate cancer (CaP; n = 55) and benign prostate hyperplasia (BPH; n = 44) specimens using immunocytochemistry (ICC) and Northern blotting. In situ hybridization (ISH), performed on a subgroup of these specimens, proved to be a more informative technique for the assessment of messenger RNA (mRNA) in this heterogeneous tissue. A comparative analysis was made in relation to the proliferative index, assessed using the MIB-1 antibody. Elevated levels of EGF-R and TGFalpha, mRNA, and protein were observed in carcinoma cells compared with benign, secretory epithelium using in situ hybridization and immunocytochemistry. In carcinoma specimens evidence of an autocrine growth loop is provided by a correlation between EGF-R and TGFalpha, mRNA (P < .0001), and protein expression (P < .01). A trend toward increased expression of EGF-R and TGFalpha protein with dedifferentiation and a similar trend in the growth fraction suggest a role in tumor progression. Although there was a correlation between EGF-R and the proliferative index (P < .01), no relationship was found between this latter parameter and TGFalpha immunoreactivity (P > .05), indicating that this growth factor may be linked with other aspects of malignant activity rather than directly stimulating proliferation.

Topics: Cell Division; Epidermal Growth Factor; ErbB Receptors; Humans; Immunohistochemistry; In Situ Hybridization; Ligands; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; RNA, Messenger; RNA, Neoplasm; Transforming Growth Factor alpha

It is widely accepted that polypeptide growth factors are involved in the growth and development of normal and neoplastic human prostate. It has been previously reported that epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I) receptors are present in the human hyperplastic prostate tissue (BPH). To add information on the mechanism of action of EGF and transforming growth factor-alpha (TGF alpha), a peptide correlated to EGF, and the EGF receptor (EGF-R) in the human prostate, we studied the expression and cellular localization of messenger ribonucleic acid (RNA) encoding EGF, EGF-R, and TGF alpha in BPH tissue. Reverse transcriptase-PCR of total RNA extracted from BPH tissues documented the presence of specific transcripts for EGF, EGF-R, and TGF alpha. In situ hybridization with specific RNA probes synthesized from the respective complementary DNA demonstrated that EGF, EGF-R, and TGF alpha messenger RNAs were mainly localized in the epithelial cells. Immunprecipitation and Western blot analysis showed that BPH tissue contained the corresponding proteins, EGF and TGF alpha. Our findings provide additional support for the idea that EGF and TGF alpha may be considered specialized symbols in the language of cell-cell interactions and for the hypothesis that in the human prostate they seem to act in an autocrine fashion.

Topics: Epidermal Growth Factor; ErbB Receptors; Female; Gene Expression; Humans; In Situ Hybridization; Kidney; Male; Placenta; Polymerase Chain Reaction; Pregnancy; Prostatic Hyperplasia; RNA, Messenger; Transforming Growth Factor alpha

Recent studies have shown that growth factors may play a role in the etiology of benign prostatic hyperplasia (BPH) and prostatic carcinoma. Several growth factors have been reported to be expressed by prostatic tissues, but these growth factors have never been examined in human fetal prostate and compared with adult prostates and cancer cell lines. The present study was designed to investigate the messenger ribonucleic acid (mRNA) expression of transforming growth factor (TGF)-alpha, TGF-beta 1, TGF-beta 2, TGF- beta 3, keratinocyte growth factor (KGF), epidermal growth factor (EGF), EGF receptor (EGF-R), and KGF receptor (KGF-R) in human fetal and adult prostatic tissues and cancer cell lines by reverse-transcriptase-polymerase chain reaction-(RT-PCR) using specific oligonucleotide primers.. Total RNA was extracted from human fetal and adult prostates (BPH tissues) and cancer cell lines. The gene expression of these growth factors and their receptors was determined by RT-PCR using specific oligonucleotide primers.. The results of these experiments suggest that: (1) human fetal prostate expressed mRNA transcripts for TGF-alpha, TGF-beta 1, TGF-beta 2, TGF-beta 3, and EGF. However, KGF, KGF-R, and EGF-R mRNA were not expressed by human fetal prostate; (2) human adult prostate (BPH tissues) showed mRNA transcripts for all growth factors and their receptors except KGF-R; (3) human BPH-1 cell lines expressed mRNA transcripts for TGF-alpha, TGF-beta 1, TGF-beta 2, TGF-beta 3, EGF, and KGF-R, but not for EGF-R and KGF growth factors; (4) human primary prostate cancer cell line (ND-1) showed mRNA transcripts for all growth factors except EGF and KGF; and (5) human prostate cancer cell lines (LNCaP, DU-145, PC-3) expressed mRNA transcripts for all growth factors except KGF, which was absent in all cell lines. However, KGF-R mRNA was absent in the PC-3 prostate cancer cell line.. These results suggest that the differential gene expression for various growth factors and their receptors in human fetal and adult prostatic tissues and cancer cell lines may be important in understanding the role of these factors in the pathophysiology of prostatic diseases.

Topics: Adult; Epidermal Growth Factor; Fetus; Fibroblast Growth Factor 10; Fibroblast Growth Factor 7; Fibroblast Growth Factors; Gene Expression; Growth Substances; Humans; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Receptors, Growth Factor; RNA, Messenger; Transforming Growth Factors; Tumor Cells, Cultured

We have established organ cultures of human prostate in vitro analysis of the hormone responsiveness of prostatic carcinoma. The influence of dihydrotestosterone (DHT), the epidermal growth factor (EGF) and prolactin (PRL) was assessed on the proliferation rate of 30 benign human prostatic hyperplasias (BPH) maintained for 48 hours as short-term primary cultures. The influence exerted by the EGF, the DHT and the PRL on the BPH cell proliferation was characterized by adding these compounds alone or in combination to the culture media. The proliferation rate was assessed by means of histo-autoradiographical nuclear labeling with titriated thymidine. The results show that of the DHT, EGF and PRL, the first two induced a statistically significant increase in the cell proliferation rate in a proportion of BPH significantly higher than the proportion of BPHs whose cell proliferation was increased significantly by the PRL. Furthermore, the PRL was able to antagonize the EGF-induced stimulatory effect on the BPH glandular cell proliferation rate. Lastly, the EGF- and PRL-mediated effects on the BPH glandular cell proliferation correlated with each other, while the DHT-mediated did not correlate with either the PRL- or the EGF-mediated effects. This model of organ, culture provide a basis for developing a method for in vitro testing of the individual hormone and drug responsiveness of a prostatic carcinoma.

Topics: Aged; Aged, 80 and over; Autoradiography; Cell Division; Dihydrotestosterone; Epidermal Growth Factor; Humans; Male; Middle Aged; Models, Biological; Organ Culture Techniques; Prolactin; Prostatic Hyperplasia; Prostatic Neoplasms

Androgens provide the primary signal for the onset of DNA synthesis and cell division in normal prostate and benign prostatic hyperplasia (BPH). It is possible, however, that androgen mitogenic activity is in part indirect and mediated by peptide growth factors. In LNCaP cell lines, R1881 added to DCC-FCS medium increases DNA, epidermal growth factor (EGF) and EGF receptor (EGFR) levels: the antiandrogen hydroxy-flutamide prevents the increase of the growth factor and increases its receptor. In BPH tissue removed by transvesical prostatectomy, DHT, testosterone, 3 alpha-androstanediol and nuclear androgen receptors (AR) show a positive linear correlation with EGF: treatment with flutamide decreases significantly the EGF production. Androgens, therefore, represent important modulatory factors of prostatic EGF release. Moreover, androgens and EGF downregulate EGFR, which is probably internalized into the cell and degraded by lisosomes: in fact, a negative linear correlation between EGF, nuclear AR and the high- and low-affinity binding of EGFR is observed. These findings support the hypothesis that the growth-promoting effects of androgens in the prostate are in part mediated by peptide growth factors. The inhibitory effect of antiandrogens on prostatic cell proliferation may be the result of the decreased androgenic support and decreased EGF release and expression.

Topics: Aged; Androstane-3,17-diol; Cell Division; Cell Line; Cell Nucleus; Dihydrotestosterone; Epidermal Growth Factor; ErbB Receptors; Gonadal Steroid Hormones; Humans; Immunohistochemistry; In Vitro Techniques; Kinetics; Male; Middle Aged; Prostate; Prostatic Hyperplasia; Receptors, Androgen; Receptors, Estrogen; Signal Transduction; Testosterone

In benign prostatic hyperplasia (BPH), basic fibroblast growth factor (bFGF) is found to have a regional distribution, with concentrations in the periurethral zone (where the primitive fibrostromal nodule originates) higher than those of the peripheral subcapsular zone. The aim of the present investigation was to verify whether androgens and epidermal growth factor (EGF) are uniformly distributed from the periurethral to the peripheral zone or whether they show regional differences. Tissue samples, removed by transvesical resection from nine untreated BPH patients, sectioned in periurethral, subcapsular, and intermediate zones, were examined. In the periurethral zone, dihydrotestosterone (DHT), testosterone, and EGF, determined by radioimmunoassay (RIA) techniques after purification on Celite microcolumns and Sep-pak C18 cartridge, showed values significantly higher (mean +/- SD: 1121 +/- 482 pg, 250 +/- 129 pg, and 6.89 +/- 3.28 ng/mg DNA, respectively; P < 0.01) than those of the subcapsular zone (489 +/- 190 pg, 114 +/- 70 pg, and 3.40 +/- 1.90 ng/mg DNA, respectively). A positive linear correlation between EGF, testosterone, and DHT was also observed. The regional distribution of EGF, testosterone, and DHT was similar to that found for bFGF: the highest levels of these factors in the periurethral region allow us to hypothesize on their possible involvement in the rewakening of mesenchymal tissue, leading to the formation of the primitive fibrostromal nodule and then to BPH development.

Topics: Data Interpretation, Statistical; Dihydrotestosterone; Epidermal Growth Factor; Fibroblast Growth Factor 2; Humans; Male; Prostate; Prostatic Hyperplasia; Radioimmunoassay; Testosterone

In order to more clearly define the status of epidermal growth factor receptor (EGFR) in prostate cancer, expression of EGFR transcript and protein was analyzed in paired samples of benign and malignant tissues from 30 radical prostatectomy specimens. Prostate tumors and high grade prostatic intraepithelial neoplasias (PINs) expressed significantly less EGFR protein than benign tissues or low grade PINs (P < 0.001). Expression of EGFR mRNA was analyzed in a subset of the same samples, and was higher in more prostate tumors than benign specimens (P < 0.05). However, differences in mean mRNA expression between malignant and benign tissues were not significant. EGFR mRNA was expressed at moderate or low levels in equivalent numbers of PIN lesions. These results suggest that, although EGFR mRNA expression is somewhat elevated in prostate tumors, EGFR protein expression may be down-regulated in the same malignant tissues. Furthermore, our data demonstrate phenotypic similarity between prostate tumors and high grade PIN at the level of EGFR protein expression.

Topics: Aged; Atrophy; Carcinoma in Situ; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization; Male; Middle Aged; Precancerous Conditions; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; RNA, Messenger; Transforming Growth Factor alpha

Enhanced expression of the epidermal growth factor receptor (EGFR) or its ligands, epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha) can increase signalling via receptor-mediated pathways which may lead to excessive proliferation and cellular transformation. Such autocrine regulation of growth has been demonstrated for prostate cancer cell lines in culture but its role in prostate cancer in vivo has not been established. To assess the potential of such a mechanism, we have examined the pathway components in prostate carcinomas (CaP) in comparison with non-malignant benign prostatic hyperplasias (BPH). In the present study, we investigate the dosage, structure and expression of EGF, TGF-alpha and EGFR genes in a series of 34 human prostate samples and 3 prostate cancer cell lines. All of the samples contained transcripts from each of the genes. The expression of pre-pro-TGF-alpha mRNA and pre-pro-EGF mRNA were significantly higher in CaP (n = 13) than BPH (n = 21) specimens (p < 0.05). The androgen-responsive prostatic carcinoma cell line, LNCaP, expressed high levels of EGF mRNA while the androgen-independent DU145 and PC-3 cell lines expressed high levels of TGF-alpha mRNA and EGFR mRNA. In general, overexpression of these mRNAs was not associated with amplification or detectable gene rearrangement; only DU145 cells demonstrated any alteration in these genes, with apparent amplification of the TGF-alpha gene. Relative to BPH, all prostate carcinomas and cell lines studied had elevated levels of mRNA for one or both mRNA coding for the ligands for EGFR.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation; Humans; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; RNA, Messenger; Transforming Growth Factor alpha; Tumor Cells, Cultured

The present immunohistochemical study examines the expression of TGF alpha, EGF, and the EGF receptor in prostatic tissues obtained from patients with either benign prostatic hyperplasia (BPH) or adenocarcinoma of the prostate. Frozen sections were cut at 5 microns and were incubated with monoclonal antibodies directed against TGF alpha, EGF, or the EGF receptor. The remainder of the staining procedure was performed with an avidin-biotin-complex peroxidase procedure. Strong TGF alpha expression was not detected in any of the 15 BPH specimens examined or in the normal/hyperplastic tissue intermixed within the adenocarcinomas. In contrast, malignant cells in 9 of 11 adenocarcinomas strongly expressed TGF alpha. EGF reactivity was observed in both hyperplastic and malignant epithelial tissues. EGF receptor expression was strongest along the basal aspects of cells of normal or hyperplastic glands. Although most malignant cells also were positive for the EGF receptor the level of staining was less than that observed in cells forming normal or hyperplastic glands. These results suggest that EGF and the EGF receptor may be components of an autocrine/paracrine regulatory pathway involved in hyperplastic and/or malignant disease of the prostate. The finding that TGF alpha is expressed only in malignant cells suggests that TGF alpha may represent a locally active autocrine regulator of malignant prostate cells.

Topics: Adenocarcinoma; Epidermal Growth Factor; ErbB Receptors; Humans; Immunohistochemistry; Male; Prostatic Hyperplasia; Prostatic Neoplasms; Staining and Labeling; Transforming Growth Factor alpha

We measured immunoreactive EGF and TGF alpha in prostate tissue extracts obtained from 19 patients with benign prostatic hyperplasia (BPH) and 19 with cancer of the prostate (CaP). Whilst both BPH and CaP expressed EGF (BPH = 195.61 +/- 19.94 ng g-1 protein; CaP = 235.60 +/- 24.45 ng g-1 protein) and TGF alpha (BPH = 92.57 +/- 7.60 ng g-1 protein; CaP = 100.73 +/- 15.47 ng g-1 protein) in equal concentrations, the levels of EGF in any tissue extract were on average twice those of TGF alpha. Furthermore analysis of the individual growth factor data revealed a direct correlation between EGF and TGF alpha in both BPH (r = 0.72, P < 0.001) and CaP (r = 0.69, P < 0.001). When the tumours were classified according to their Gleason score, a slight but significant increase in growth factor concentrations was noted as the tumour became less differentiated. We also measured the concentrations of testosterone and dihydrotestosterone (DHT) in prostate extracts with a view of elucidating the relationship between androgen and growth factors in this gland. There was a small positive correlation only between testosterone and EGF (r = 0.62, P < 0.05) and testosterone and TGF alpha (r = 0.61, P < 0.05) in CaP. The absence of any similar correlation in BPH where DHT becomes the predominant hormone may suggest an indirect role for testosterone in the regulation of growth factor production.

Topics: Androgens; Dihydrotestosterone; Epidermal Growth Factor; Humans; Male; Prostatic Hyperplasia; Prostatic Neoplasms; Testosterone; Transforming Growth Factor alpha

Human benign prostatic hyperplasia (BPH) samples were analyzed to evaluate the presence of immunoreactive epidermal growth factor (irEGF) and EGF receptor (EGFR). In all BPH samples examined both peptide and its receptor were present. Scatchard analysis of binding data of [125I]EGF showed two classes of binding sites with high and low affinity. Intratissular irEGF concentrations showed a significant inverse linear correlation with EGFR levels. Two groups of samples can be identified: the first showing high irEGF concentrations and low levels of EGF binding sites; the second low irEGF and high concentrations of EGFR. The simultaneous presence of EGF and its receptor in BPH samples indicates that this growth factor may act in an autocrine/paracrine manner in human prostatic tissue. The inverse relationship between EGF and the two sites of EGFR lead one to hypothesize that EGF itself could play a central role in determining receptor cell surface availability.

Topics: Aged; Cell Membrane; Epidermal Growth Factor; ErbB Receptors; Humans; Immunoassay; Male; Middle Aged; Prostatic Hyperplasia

Stromal enlargement plays a key role in the development of benign prostatic hypertrophy in humans. Human prostatic fibroblasts were obtained from fetal and adult prostates and characterized as to their androgen and estrogen receptor status and growth in response to dihydrotestosterone (DHT), estradiol (E2), hydroxyflutamide (OH-FLU), hydrocortisone, basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF). In addition, the ability of hormones and growth factors to induce the messenger RNA (mRNA) for the c-fos protooncogene was assessed as a measure of the early, direct effects of these compounds on cellular proliferation. Nuclear androgen receptors were demonstrable by immunocytochemistry in both fetal and adult cells. Nuclear estrogen receptor staining was negative. Neither E2 nor hydrocortisone increased cellular proliferation. Both EGF and bFGF did increase cellular growth. DHT (10(-8)-10(-7) M) had a significant stimulatory effect on cell growth only in serum-free media. OH-FLU addition enhanced DHT induced proliferation. Changing the media during the course of the experiment obliterated the stimulatory effect of DHT. Both EGF (10 ng/ml) and bFGF (20 ng/ml) increased the mRNA for the c-fos protooncogene. DHT (10(-7) M) did not induce the mRNA for c-fos. We conclude that EGF, bFGF, and DHT (especially in combination with OH-FLU) increase the proliferation of human prostatic fetal and adult fibroblasts in vitro. E2 has no effect on fibroblast proliferation. The stimulatory effects of EGF and bFGF are direct, whereas the effect of DHT appears to be indirect, possibly mediated via the increased production and/or secretion of growth factors.

Topics: Cell Division; Cells, Cultured; Dihydrotestosterone; Epidermal Growth Factor; Estradiol; Female; Fetus; Fibroblasts; Flutamide; Genes, fos; Growth Substances; Humans; Immunohistochemistry; Male; Pregnancy; Prostate; Prostatic Hyperplasia; RNA, Messenger

Benign prostatic hyperplasia (BPH) is a sex steroid dependent disease. Estrogens and androgens can modulate in different mammalian tissues epidermal growth factor (EGF) production and/or secretion. In order to clarify the relationships between estrogen and androgen receptor concentrations and those of immunoreactive EGF (irEGF), we have evaluated these parameters in 14 human BPH samples, by means of a dextran-coated charcoal method and radioimmunoassay, respectively. Cytosolic steroid receptors did not seem to correlate with irEGF. A linear significative relationship was evident between nuclear androgen receptor (ARn) levels and endogenous irEGF but not between nuclear estrogen receptors and irEGF: in ARn negative BPH samples, irEGF levels were lower than in ARn positive ones. Therefore, it is possible that androgens act at prostatic tissue level, through their own receptors, by modulating EGF production and/or secretion.

Topics: Aged; Cell Nucleus; Cytosol; DNA; Epidermal Growth Factor; Humans; Male; Middle Aged; Prostate; Prostatic Hyperplasia; Radioimmunoassay; Receptors, Androgen; Receptors, Estrogen

To evaluate the distribution and density of epidermal growth factor (EGF) receptors (EGF-Rs) on urothelium, immunohistological studies using a monoclonal antibody to the binding portion of the human EGF-R were performed on frozen specimens of normal urothelium (N = 20), urothelium from patients with nonurothelial urological malignancies (N = 15) and inflammatory diseases (N = 8), low grade superficial transitional cell carcinomas (TCC) (N = 13), high grade superficial or invasive TCC (N = 28), and endoscopically normal appearing urothelium from patients with low grade superficial (N = 5) or high grade (N = 21) TCC elsewhere in the bladder (or ipsilateral renal pelvis/ureter). EGF-Rs are found only on the basal layer of epithelial cells (with scattered representation on intermediate cells) in 95% of normal urothelial specimens and 100% of pathological specimens without urothelial malignancy. Alternatively, 92.3% of specimens of low grade superficial TCC and 100% of high grade TCCs had EGF-Rs richly expressed on the superficial as well as the deeper layers of urothelium. This "malignant" distribution of EGF-Rs was also found on all specimens of endoscopically normal appearing urothelium in patients with TCC elsewhere. The density of EGF-Rs correlated closely with tumor grade on both "premalignant" and frankly neoplastic urothelium. We conclude that the expression of EGF-Rs on urothelium favors the interaction of premalignant and malignant tissue with urinary EGF. To determine if altering the physiochemical environment of urine could interfere with this interaction, the effects of pH on the binding of and growth responses to EGF were assessed on four human TCC cell lines. Scatchard plots demonstrated that varying pH from 5.0 to 7.5 did not significantly change the total number of receptors, but EGF-R affinity was reduced approximately 20-fold as pH decreased from 7.5 to 5 in each TCC target. Similarly, significant growth stimulation by EGF at pH 7.5 was abrogated at pH less than or equal to 7.0 while growth rates in the absence of EGF remained unchanged at lower pHs. It thus appears that urinary acidification may hold promise in the management and prevention of recurrent bladder cancer.

Topics: Adenocarcinoma; Carcinoma, Transitional Cell; Cell Division; Epidermal Growth Factor; Epithelium; ErbB Receptors; Female; Humans; Kinetics; Male; Neoplasm Invasiveness; Prostatic Hyperplasia; Prostatic Neoplasms; Tumor Cells, Cultured; Ureter; Urinary Bladder; Urinary Bladder Neoplasms

We characterized the epidermal growth factor (EGF) receptor in the membrane fraction of prostatic tissue from men with benign prostatic hyperplasia (BPH). The maximum specific binding of [125I]EGF to the BPH membrane fraction was achieved after 30-min incubation at 35 C. Analysis of the binding data revealed two classes of binding sites, one of high affinity [Kd, 2.5 +/- 0.5 (+/- SE) x 10(-11) mol/L] and one of lower affinity (2.2 +/- 0.3 x 10(-9) mol/L). [125I]EGF binding was inhibited by excess EGF, but not by insulin, proinsulin, fibroblast growth factor, or insulin-like growth factors I and II. In prostatic tissue of men with BPH treated for 3 months with the GnRH agonist analog Goserelin (Zoladex, depot formulation), the binding capacities of both sites were significantly higher than those of BPH tissue from untreated men (P less than 0.001). These results demonstrate that prostatic tissue from men with BPH contains two classes of specific binding sites for EGF, and their levels are modulated by chronic GnRH agonists treatment.

Topics: Aged; Binding Sites; Buserelin; Dihydrotestosterone; Epidermal Growth Factor; ErbB Receptors; Goserelin; Humans; Male; Middle Aged; Models, Biological; Prostatic Hyperplasia; Testosterone

The receptor for epidermal growth factor (EGF-R) was characterized on membrane fractions from human benign prostatic hyperplasia (BPH). Specific binding of [125I]EGF reached equilibrium after 40 min at 25 degrees C and was stable for up to 120 min. Saturation analysis of EGF-R, performed by incubating the membranes with 0.0156-15 nM [125I]EGF in the presence and in the absence of 100-fold excess of cold EGF for 60 min, revealed the presence of two classes of binding sites with high and low affinities (Kd = 0.35 +/- 0.23 and 9.60 +/- 2.87 nM respectively). Competition experiments revealed that FSH, insulin and calcitonin did not compete with [125I]EGF. The simultaneous determination of EGF-R and that of estradiol (ER), progesterone (PR) and androgen receptors (AR) was performed using the same buffer to homogenate the tissues and to obtain cellular membranes. The steroid receptors (SR) were determined by means of the dextran-coated charcoal method. There was a significant negative correlation between nuclear SR and binding capacity of EGF-R. The presence of specific and high affinity binding sites for EGF and the modulation of the level of these sites by steroid receptors suggest a possible role of EGF in prostatic hyperplasia.

Topics: Biomarkers; Cell Nucleus; Epidermal Growth Factor; ErbB Receptors; Humans; Kinetics; Male; Prostate; Prostatic Hyperplasia; Receptors, Steroid

Associations between epidermal growth factor (EGF) and carcinoma of the prostate (CAP) have not been systematically investigated. We used indirect immunohistochemical techniques to demonstrate cytoplasmic EGF in paraffin-embedded sections of the following primary prostatic tissues: benign prostatic hyperplasia (BPH) (N = 10), BPH adjacent to CAP (N = 42), clinically localized CAP (N = 45), untreated metastatic CAP (N = 10), and metastatic CAP after varying periods of androgen deprivation (N = 10). In six of the latter 10 cases biopsies of the primary tumor obtained before androgen deprivation therapy were also available for study. Three of the BPH specimens (6%) and 44 of the CAP specimens (68%) stained. Forty per cent of the localized tumors stained but all untreated and treated metastatic tumors stained (p less than 0.01). There were direct but statistically insignificant correlations between the demonstration of EGF and both the Gleason score of localized and untreated metastatic tumors and the pathologic stage of localized tumors. The proportion of malignant cells stained in EGF positive tumors was similar regardless of Gleason score, pathologic stage or the presence or absence of metastases. However, the proportion of cells stained was greater in five of six specimens obtained during hormonal deprivation compared to specimens of the same tumor obtained before treatment. These data suggest that some prostatic cancers interact with EGF and that the interaction may be influenced by the androgenic milieu.

Topics: Adenocarcinoma; Epidermal Growth Factor; Humans; Immunoenzyme Techniques; Male; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms

High concentrations (272 +/- 33 ng/ml) of urogastrone-epidermal growth factor were measured in prostatic fluid from normal males by a specific radioimmunoassay. Significantly lower concentrations (155 +/- 24 ng/ml) were observed in the prostatic fluid of patients with benign prostatic hypertrophy than in the age-matched normal controls (2P less than 0.01). The growth factor content of seminal fluid was accounted for by the contribution of prostatic fluid. Immunochemical studies failed to show evidence of synthesis within the gland nor could high affinity receptors for the protein be demonstrated in membrane preparations of the gland.

Topics: Chromatography, Gel; Citrates; Citric Acid; Epidermal Growth Factor; Humans; Male; Phosphates; Prostate; Prostatic Hyperplasia; Zinc

A growth factor capable of stimulating DNA synthesis of BALB/3T3 cells was purified about 1,000-fold from the cytosol of human benign hypertrophic prostates by heparin-Sepharose chromatography; the growth factor bound to the column in the presence of 0.5 M NaCl was eluted with 1.5-1.7 M NaCl. Its molecular weight and isoelectric point were estimated to be 11,000-13,000 and 10.5, respectively. It was sensitive to heat- and acid-treatments but resistant to disulfide-reducing agent. The final preparation was able to stimulate DNA synthesis at 10 ng/ml. The degree of stimulation was dependent on serum concentration in the assay system; the degree of maximum stimulation increased about 5 times as serum concentration increased from 0.2 to 2%.

Topics: Cytosol; Epidermal Growth Factor; Growth Substances; Humans; Isoelectric Point; Male; Molecular Weight; Prostate; Prostatic Hyperplasia

Extracts of benign prostatic hyperplasia (BPH) contain a factor which is mitogenic for human foreskin fibroblasts in culture. Because of the similarity of BPH extract and epidermal growth factor (EGF) in stimulating quiescent fibroblasts to divide, it was of interest to determine if the prostate-associated growth factor competes for EGF receptor binding. BPH extract was found to compete poorly for 125I-EGF-receptor binding, did not influence the dissociation of cell-bound 125I-EGF and caused only a slight down-regulation of the EGF receptor. These findings indicate that EGF and BPH extract do not recognize the same receptor and that the major growth stimulating activity of BPH extract is not due to EGF.

Topics: Binding, Competitive; Epidermal Growth Factor; Fibroblasts; Humans; Male; Mitosis; Prostate; Prostatic Hyperplasia; Tissue Extracts

Topics: Adenocarcinoma; Cell Aggregation; Cell Line; Cell Survival; Culture Media; Epidermal Growth Factor; Epithelial Cells; Humans; Karyotyping; Male; Microscopy, Electron, Scanning; Microvilli; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms