bluuebunny/crossref_metadata_2025 huggingface dataset:
Androgen ablation induces tenascin expression in the rat prostate
Authors: Vollmer Günter , Michna Horst , Ebert Kirsten , Knuppen Rudolf
Published: February 2007

Tenascin is a glycoprotein of the extracellular matrix of mesenchymal derived tissue compartments. Although the DNA sequences that code for tenascin production are known and the protein structure is well characterized, little is known about regulation of tenascin expression. Therefore, we are interested in hormonal aspects of tenascin expression. In this study, we addressed the question if androgen deprivation and prostatic involution would influence tenascin expression in the prostate. Methodologically, in two series of experiments, intact and orchiectomized testosterone propionate substituted male rats were subjected to one of the following hormonal treatments: a) flutamide, b) the antiandrogen casodex, and c) cyproterone acetate (CPA). As controls in each series, we used untreated controls and orchiectomized rats. After a period of 14 days of treatment, prostates were removed. Tenascin immunostaining of the sectioned specimen from the control and the hormonally treated animals demonstrated the following: 1) Little, if any tenascin immunoreactivity was detectable in prostates of untreated animals. 2) Androgen deprivation with either treatment resulted in tenascin expression in the stroma of the prostates. 3) Tenascin expression appeared to be variable both semiquantitatively and in the staining pattern detectable except in prostates treated with CPA, in which we observed the most uniform and most widespread staining pattern. From these results, we conclude that androgen deprivation induces tenascin expression in the stroma of the involuting prostate. Remodelling of the extracellular matrix of the stroma, known to appear in the process of prostate involution by androgen ablation, represents a process which has not been discussed with tenascin expression so far. © 1994 Wiley‐Liss, Inc.
Detection of prostate cancer in males with prostatism
Authors: Lepor Herbert , Owens R. Scott , Rogenes Vince , Kuhn Evelyn
Published: February 2007

The present study was designed to compare the prostate cancer detection rate, sensitivity, specificity, and positive predictive value of digital rectal examination (DRE) and serum prostatic specific antigen (PSA) in a consecutive cohort of males presenting to a single institution with clinically significant prostatism. The study population was comprised of 224 consecutive males with clinically significant prostatism referred to the Prostate Center at the Medical College of Wisconsin between June 1990 and December 1991. Subjects were considered to have clinically significant prostatism if they elected to pursue medical or surgical therapy following exclusion of carcinoma of the prostate.

The initial examination consisted of a Boyarsky symptom score assessment, DRE, uroflowmetry, postvoid residual determination, serum PSA level, and transrectal prostatic ultrasonography. Subjects with an abnormality on DRE or serum PSA > 4 ng/dl were advised to undergo transrectal prostatic biopsy. Of the 224 subjects, 40 (17.9%) had an abnormal DRE and 57 (25.4%) had an elevated serum PSA > 4 ng/dl. The overall detection rate of prostate cancer in the study population was 6.7%. The prostate cancer detection rates for PSA alone and DRE alone were 5.8% and 5.3%, respectively. The sensitivity, specificity, and positive predictive values of PSA alone were 86.7%, 80.9%, and 25.0% and of DRE alone 80.0%, 86.3%, and 30.0%, respectively.

Receiver operator characteristic (ROC) curves were constructed for the entire study population in order to compare the screening measures serum PSA and PSA density. The area under the curves was 0.88 for both tests, indicating that these screening tests for prostate cancer were not significantly different. The present study demonstrated that males with clinically significant prostatism represent a high risk cohort for detecting prostate cancer. DRE and PSA are equally effective measures for detecting prostate cancer. PSA density does not offer any advantage over serum PSA in screening for prostate cancer, except in the subset of patients with a normal DRE and serum PSA levels between 4.0 and 9.9 ng/dl. © 1994 Wiley‐Liss, Inc.
Pattern of prostate cancer metastasis to the vertebral column
Authors: Suzuki Takanori , Shimizu Toshihiro , Kurokawa Kohei , Jimbo Hiroyuki , Sato Jin , Yamanaka Hidetoshi
Published: February 2007

We examined 34 thoracic and lumbar vertebrae in eight patients with prostate cancer to elucidate the pattern of metastasis to the vertebral column. Computed tomography of thoracic and lumbar vertebrae was performed before treatment for the prostate cancer. Metastatic lesions were confirmed both by the recognition of osteoblastic and/or osteolytic lesions on CT and by histological identification at autopsy. Localized metastatic lesions were observed mostly in the peripheral rather than the central part of the vertebral body. There were no differences in distribution of lesions between the anterior and posterior parts of the vertebral body. There was no primary involvement of tumor in the spinous and transverse processes. These findings indicated that the major metastatic pathway to the vertebral column is via the vertebral venous system, and it appears that prostate cancer cells first metastasize to the vertebral body and then spread secondarily to the processes or other vertebrae. © 1994 Wiley‐Liss, Inc.
Histological evaluation of benign prostatic hyperplasia treated by long‐term administration of chlormadinone acetate (CMA)
Authors: Harada Masaoki , Kinoshita Yuzo , Moriyama Masatoshi , Kondo Iichiro , Nakahashi Mitsuru , Kumagai Harumi , Sasaki Koichi , Hosaka Masahiko
Published: February 2007

Although the clinical effects of attempted nonsurgical treatment of benign prostatic hyperplasia have been well documented, detailed histological evaluation of the effects of treatment appears to be limited. The effect of long‐term administration of an antiandrogen, chlormadinone acetate (CMA), on benign prostatic hyperplasia was evaluated with histological comparison of two biopsy specimens, one before treatment and one after treatment. Secretory epithelium showed obvious regressive changes with occasional basal cell prominence after CMA treatment. Stromal elements, however, did not show any marked changes, except for occasional edematous loosening. Scores of multiple epithelial parameters tended to be correlated with clinical improvement in urinary obstructive symptoms, especially in patients with predominant glandular hyperplasia. These results suggest that long‐term administration of the potent antiandrogen CMA to inhibit dihydrotestosterone‐receptor binding might be a useful therapeutic maneuver in patients with glandular hyperplasia, without any deterioration of the stromal component. © 1994 Wiley‐Liss, Inc.
Synergistic effects of estrogen with androgen on the prostate—effects of estrogen on the prostate of androgen‐administered rats and 5‐alpha‐reductase activity
Authors: Suzuki Kazuhiro , Takezawa Yutaka , Suzuki Takanori , Honma Seijiro , Yamanaka Hidetoshi
Published: February 2007

To clarify the effects of estrogen on the prostate of androgen‐treated rats, we administered estradiol (E2; 0.01 mg/day) to testosterone (T; 1 mg/day)‐treated Wistar rats under various conditions. There were three variable factors: the age of the rat, the untreated period after castration, and the presence of testes. Under all conditions, E2 stimulated the growth of the prostate in T‐treated rats compared with growth in rats in treated with T only.

To analyze the mechanism underlying this enhancement of prostate growth by estrogen, we measured 5‐alpha‐reductase activity and calculated its kinetic parameters, i.e., both Vmax and Km. The Vmax of the nuclear fraction was higher in E2‐administered T‐treated rats than in T only‐treated rats. In contrast, the Vmax of the microsomal fraction was lower in E2‐administered rats. Km values in the two groups showed no significant differences. Elevation of the nuclear fraction of 5‐alpha‐reductase activity could explain the synergistic effects of estrogen on the prostate growth of androgen‐treated rats. © 1994 Wiley‐Liss, Inc.
In vivo and in vitro effects of androgen on fibroblast growth factor‐2 concentrations in the human prostate
Authors: Geller Jack , Sionit Lida R. , Baird Andrew , Kohls Matthew , Connors Kenneth M. , Hoffman Robert M.
Published: February 2007

Prostatic growth is primarily regulated by dihydrotestosterone (DHT). Recent studies have demonstrated that a large number of growth factors are present in the human benign prostatic hyperplasia (BPH) prostate, including epidermal growth factor (EGF), transforming growth factor‐alpha (TGF‐α), transforming growth factor beta (TGF‐β), insulin‐like growth factor (IGF), and basic fibroblast growth factor (bFGF) (FGF‐2). DHT may mediate its mitogenic effects in the prostate by regulating growth factors. To test this hypothesis, we have utilized a histoculture androgen sensitivity assay (HASA) in which 3H‐thymidine incorporation is measured in aliquots of BPH tissue in histoculture with either added DHT or hydroxyflutamide (HF). The resulting DHT/HF ratio is an expression of the androgen sensitivity of the tissue. In this study, we have compared the DHT/HF ratio for 3H‐thymidine incorporation to the DHT/HF ratio for FGF‐2 measured in the histocultured prostates. The DHT/HF ratio for the HASA studies of 3H‐thymidine incorporation averaged 2.68 compared to the DHT/HF ratio for FGF‐2 in the same specimens of 1.01. These values were significantly different, therefore indicating no relationship between DHT stimulation and FGF‐2 levels. In addition, FGF‐2 levels were measured in human BPH prostates from patients medically castrated with megesterol acetate and estradiol 17‐β prior to surgery. These values were not significantly different, and therefore do not suggest any effect of DHT on the concentration of prostatic FGF‐2. Although these studies did not show any effect of DHT on the regulation of prostatic FGF‐2, they do indicate that the HASA assay is feasible and appropriate to use in the study of relationships between DHT and various growth factors. © 1994 Wiley‐Liss, Inc.
Transforming growth factor beta I expression in benign and malignant prostatic tumors
Authors: Glynne‐Jones E. , Harper M. E. , Goddard L. , Eaton C. L. , Matthews P. N. , Griffiths K.
Published: February 2007

The expression of transforming growth factor beta 1 (TGF‐β1) in prostate specimens obtained from patients with benign prostatic hyperplasia (BPH, n = 32) and prostate carcinoma (n = 66) was investigated using Northern blot analysis and immunohistochemistry. Northern blot analysis revealed TGF‐β1 message (2.5 kb) in virtually all of the samples examined, reflecting the ubiquitous nature of this growth factor. No statistical difference was found between the levels of mRNA detected in benign and malignant tissues due, in part, to the inherent heterogeneity of prostate tissue. Immunohistochemical methods using an antibody to native TGF‐β1 revealed a novel pattern of immunoreactivity. Staining observed only in certain epithelial cells of benign glands was associated with areas of infection rather than tumorigenesis. Interestingly, intense staining was also seen in polymorphonuclear leukocytes. No correlation was found with the mRNA results, suggesting that this antibody is binding to TGF‐β1 activated in response to infection rather than detecting sites of synthesis of latent TGF‐β. © 1994 Wiley‐Liss, Inc.
Loss and gain of chromosomes 1, 18, and Y in prostate cancer
Authors: König Josée J. , Teubel Wilma , Van Dongen Jan Willem , Romijn Johannes C. , Hagemeijer Anne , Schröder Fritz H.
Published: February 2007

Nuclear suspensions of 42 prostate carcinoma specimens obtained at surgery were used to investigate loss and gain of chromosomes 1, 18, and Y by fluorescence in situ hybridization (FISH) with centromere‐specific probes. The outcome of FISH analysis was correlated with clinical parameters and the relationship between DNA‐FCM (ploidy at cellular level) and FISH (ploidy of individual chromosomes) was assessed.

Significant loss of chromosomes 1 and 18 was infrequent (respectively, three and five cases), but 53% of the tested specimens showed loss of Y. Loss was not correlated with DNA ploidy. Significant gain occurred in 36% (chromosome 1), 63% (chromosome 18), and 28% (Y) of the specimens. Gain of chromosome 18 was shown in DNA diploid (7/14) and aneuploid tumors (18/26), while gain of chromosomes 1 and Y was nearly restricted to DNA aneuploid specimens. Significant unbalance between these chromosomes occurred in 11 cases. Most cases which had significant gain of chromosome 1 or 18 showed trisomic as well as tetrasomic cells. Simultaneous loss of some and gain of other investigated chromosomes is suggestive of clonal heterogeneity and/or multiclonality. This was observed in eight tumors.

Correlation between DNA‐FCM and FISH was best for the Y chromosome. DNA‐FCM showed more aberrant histograms with increasing stage and grade of tumors. The presence of numerical aberrations of the investigated chromosomes however, seemed independent of clinical grade or stage. © 1994 Wiley‐Liss, Inc.
Inhibition of growth and increase of acid phosphatase by testosterone on androgen‐independent murine prostatic cancer cells transfected with androgen receptor cDNA
Authors: Suzuki Hiroyoshi , Nihei Naoki , Sato Naohide , Ichikawa Tomohiko , Mizokami Atsushi , Shimazaki Jun
Published: February 2007

Most androgen‐unresponsive prostatic cancer cells are found to lack androgen receptor (AR). To clarify the role of AR in the process of the progression from androgen‐dependent to androgen‐unresponsive tumor, the AR gene was transfected into an AR‐negative rat prostatic cancer cell line CUB‐II. AR‐transfectant cells expressed AR mRNA and showed binding to R1881. AR was found in nuclei of AR‐transfectant cells by histochemical examination. Therefore, AR‐transfectant cells were considered to contain functional AR. The growth of AR‐transfectant cells was markedly inhibited in culture in the presence of testosterone, and the effect of testosterone was reduced by simultaneous addition of flutamide. Moreover, tumors inoculated with AR‐transfectant cells in male mice showed much slower growth than those in females. The tumors of AR‐transfectant cells in mice consisted of slightly larger spindle‐shaped cells when compared to those of CUB‐II cells. Moreover, AR‐transfectant cells contained a few polynuclear giant cells. Since CUB‐II cells contained acid phosphatase (AcP) activity, the addition of testosterone in culture increased AcP activity of AR‐transfectant cells. It is concluded that resumption of androgen‐dependent processes reduces the growth rate accompanying changes of phenotype. © 1994 Wiley‐Liss, Inc.
Analysis of prostatic fluid: Evidence for the presence of a prospective marker for prostatic cancer
Authors: Grover Phulwinder K. , Resnick Martin I.
Published: February 2007

In an endeavor to identify marker(s) for prostatic cancer, proteins in prostatic fluids were analyzed by two‐dimensional (2‐D) gel electrophoresis. The fluids were obtained from five males who had no prostate lesions and five patients each with benign prostatic hyperplasia (BPH) and prostatic carcinoma (PCA). The specimens were collected directly over a mixture of protease inhibitors and centrifuged, and the supernatants were lyophilized and solubilized in sodium dodecyl sulfate mix. Identical amounts of proteins were pooled according to donors' prostate disease and the resulting samples were subjected to 2‐D gel analysis employing the ISO‐DALT system. The electrophoretograms were developed by silver or double stain.

The samples of each group exhibited distinctive profiles with the exception of similar relative positions of major protein spots. A predominant protein occurring as several charge variants was consistently present in prostatic fluids of patients with PCA. This protein appeared to be a previously unknown constituent that we have called protein D (molecular weight ∼︁22 kDa and isoelectric point ∼︁4), and was undetectable in the fluids of “normal” men and patients with BPH. An analysis of pooled, unprocessed urine from PCA patients revealed that perhaps this protein is excreted in urine in very low quantities.

These results strongly suggest that the potential of this protein as a marker for prostatic cancer should be further explored.
Effects of prostatectomy on testicular androgenesis and serum levels of gonadotropins in mature albino rats
Authors: Ghosh Pradip Kumar , Ghosh D. , Ghosh P. , Chaudhuri A.
Published: February 2007

The effects of prostatectomy on testicular steroidogenic enzymes, and on serum levels of gonadotropins, prolactin, and testosterone were studied. Adult male rats were prostatectomized and sacrificed after 14 and 21 days. There was augmentation of both Δ5‐3β‐hydroxysteroid dehydrogenase (Δ5‐3β‐HSD) and 17β‐hydroxysteroid dehydrogenase (17β‐HSD) activities in the testes along with increased levels of FSH, prolactin, and testosterone in the serum, while no changes were observed in serum levels of LH. Hence it may be concluded that the prostate gland has an inhibitory effect on testicular androgenesis and can exert some influence in the regulation of FSH and prolactin secretion.
6‐methylene progesterone is cytotoxic to human cancer cell lines independent of its 5‐α‐reductase activity
Authors: Roberson Kathryn M. , Robertson Cary N. , Padilla George M. , Schmidt James Malcolm , Petrow Vladimir
Published: February 2007

This investigation examined the effects of 6‐methylene progesterone (6MP), an irreversible inhibitor of 5‐α‐reductase, on prostatic cancer (PC) cell lines. Dose titration microculture tetrazolium assays were used to evaluate cytotoxicity in cultures treated for 72 hr with 6MP (0–20 μg/ml). An androgen‐sensitive cell line, LNCaP, was drug‐sensitive with a mean 50% lethal dose value (LD50) of 2.632 ± 0.103. Hormone‐resistant PC cell lines 1‐LN, DU 145, and PC3 also demonstrated sensitivity with LD50 values between 0.8579–1.110 μg/ml with a group average of 1.023 ± 0.082 μg/ml. Increasing dosages of dihydrotestosterone in the growth media did not alter 6MP cytotoxicity in androgen‐insensitive prostatic cancer cell lines. No correlation between androgen‐responsiveness and 6MP‐induced cytotoxicity was observed. In nonprostatic malignancies, 6MP inhibited adenocarcinoma cell lines with a mean group LD50 value of 0.7772 μg/ml ± 0.110. J82, a transitional cell carcinoma cell line of bladder origin, exhibited an average LD50 value of 1.041 ± 0.260. In an epidermoid cervical cancer cell line, ME180, an LD50 value of 0.5356 μg/ml ± 0.010 was noted. In a melanoma cell line, Du Mel 6, a mean LD50 of 0.7428 ± 0.023 μg/ml was achieved with 6MP. We conclude that 6MP, a novel 5‐α‐reductase inhibitor, has potential as a cytotoxic agent in prostatic carcinoma and additional human malignancies. Further study is justified.
Allelic loss of the retinoblastoma gene in primary human prostatic adenocarcinomas
Authors: Brooks James D. , Steven Bova G. , Isaacs William B.
Published: February 2007

Inactivation of the retinoblastoma (Rb) gene has been implicated in the genesis and progression of a number of tumor types, including prostatic adenocarcinomas. We have analyzed a series of 46 surgically‐resected human prostatic adenocarcinomas for allelic loss of the Rb gene with PCR amplification of a highly polymorphic region of the gene. 41 of 46 tumors (89%) were informative and 11 of these (27%) had lost one Rb allele. The relative frequency of this occurrence suggests that inactivation of the retinoblastoma gene may be an important event in prostate carcinogenesis.
A prospective study of plasma hormone levels, nonhormonal factors, and development of benign prostatic hyperplasia
Authors: Gann Peter H. , Hennekens Charles H. , Grodstein Francine , Stampfer Meir J. , Longcope Christopher , Verhoek‐Oftedahl Wendy
Published: February 2007

We assessed the relation of plasma hormone levels and nonhormonal factors with subsequent occurrence of surgical treatment for benign prostatic hyperplasia (BPH) among participants in the Physicians' Health Study. Frozen plasma samples, collected at the study onset, were available for 320 men who developed surgically treated BPH up to 9 years later and for 320 age‐matched controls. Plasma testosterone (T), dihydrotestosterone (DHT), androstenedione, estradiol (E2), and estrone (E1) were measured for each case‐control pair. In unadjusted analyses, none of the hormones or hormone ratios were associated with BPH; for example, for T and E2 the odds ratios (OR) comparing the highest quintile (Q5) with the lowest (Q1) were 0.74 (95% CI = 0.42, 1.30) and 1.07 (95% CI = 0.51, 2.22), respectively. However, in multivariate analyses controlling diastolic blood pressure, exercise, alcohol, E1, and DHT:T ratio, we observed a strong trend for increasing risk across quintiles for E2 (Q5 vs. Q1 OR = 3.56, P trend = 0.009), and a weak inverse trend for E1 (Q5 vs Q1 OR = 0.51, P trend = 0.07). The excess risk associated with E2 was confined to men with relatively low androgen levels. Three nonhormonal factors previously suspected as risk factors were independently associated with surgical BPH in these data. The OR for a 1‐mm Hg difference in diastolic blood pressure was 1.04 (95% CI = 1.01, 1.07). Alcohol use and infrequent exercise were inversely associated with risk of BPH surgery; however, risk estimates were not consistent across categories of exercise and alcohol frequency. Our results indicate that normal variation in circulating androgen levels does not influence development of BPH, but that variation in estrogen levels might be important.
Comparison of the effects of new specific azasteroid inhibitors of steroid 5α‐reductase on canine hyperplastic prostate: Suppression of prostatic DHT correlated with prostate regression
Authors: Cohen S. M. , Werrmann J. G. , Rasmusson G. H. , Tanaka W. K. , Malatesta P. F. , Harris G. , Prahalada S. , Jacobs J. G. , Nett T. M.
Published: February 2007

Four new azasteroid inhibitors of steroid 5α‐reductase were compared to the benchmark compound finasteride, each at a dose level of 1 mg/kg/day, as well to placebo and to castration, in seven groups of mature male beagle dogs with enlarged prostates. Prostate volumes were measured repetitively by a volume MRI method over 15 weeks of treatment. The study probed the obverse of the familiar relation between DHT and prostate growth, and provides the first documentation of a tight negative correlation between prostate regression and the prostatic concentration of DHT across a range of treatment regimens (r = −0.982). In this first direct comparison study of structure vs. in vivo activity for several azasteroids in the dog model of BPH, relative efficacy for induction of shrinkage of the dog prostate did not correlate at all with the inhibitor's relative activity against the dog 5α‐reductase in vitro. On the basis of the relative IC50 values it would not have been predicted that, at the dose tested, the analogue MK‐434 (17β‐benzoyl‐4‐aza‐5α‐androst‐1‐en‐3‐one) was distinguished from the other inhibitors with respect to the induction of faster and more complete regression (69%) as well as greater reduction in prostatic DHT (95%), both of which approached the castrated dog levels of 75% prostatic shrinkage and >98% reduction in DHT. Treatment with any one of the five azasteroids induced two‐ to five‐fold increases in prostatic testosterone. However, total androgen was conserved at the placebo control level. Despite the differences noted, each azasteroid tested induced a highly significant decrease in prostatic volume that correlated tightly with a decreased prostatic DHT level in canine spontaneous BPH.
Cellular proliferation in prostatic adenocarcinoma as assessed by bromodeoxyuridine uptake and Ki‐67 and PCNA expression
Authors: Cher Michael L. , Carroll Peter R. , Chew Karen , Rosenau Werner
Published: February 2007

Prostatic carcinomas vary in their biological potential, even when stratified by grade and stage. Measurement of cellular proliferation by various methods has been shown to correlate with outcome for several human cancers, including prostatic carcinoma. Uptake of bromodeoxyuridine (BrdUrd), a thymidine analogue, has been accepted as a measure of cellular proliferative rate. However, the technique is somewhat complex, requiring incubation with fresh tissue. We compared cellular proliferation as measured by BrdUrd uptake with two more simple immunohistochemical methods in 44 prostatic adenocarcinoma specimens and correlated the results with standard clinical parameters. The tissue was obtained via needle biopsy, channel transurethral resection, and radical prostatectomy. Specimens were incubated in vitro with BrdUrd and then fixed and paraffin embedded. Sections were immunohistochemically stained with antibodies to BrdUrd, proliferating cell nuclear antigen (PCNA), and Ki‐67. At least 1,000 cells were scored, and a labeling index (LI) was calculated (number of positive cells/total number of cells). The mean LI determined by all three indices was low (BrdUrd = 3.0, PCNA = 7.0, Ki‐67 = 3.4), consistent with the knowledge that prostatic tumors grow slowly. In 36 patients who had not been treated at the time of analysis, the LI as determined by all three methods correlated well with clinical stage and pathological grade. Furthermore, the LIs discriminated between those with tumor confined to the prostate and those with extension to the seminal vesicles, lymph nodes, or bone (P = 0.003, 0.004, 0.008 for BrdUrd, PCNA, and Ki‐67, respectively). The LIs for PCNA and Ki‐67 correlated well with that for BrdUrd (r = 0.84; r = 0.85), while the LIs for Ki‐67 and PCNA correlated slightly less well with each other (r = 0.78). PCNA and Ki‐67 expression appear to reflect essentially the same biological process as BrdUrd uptake. Either can substitute for BrdUrd as a measure of cellular proliferation, and Ki‐67 seems to offer the fewest technical problems.
Differential expression of uPA in an aggressive (DU 145) and a nonaggressive (1013L) human prostate cancer xenograft
Authors: Billström Anita , Dahllöf Björn , Hartley‐Asp Beryl , Lecander Ingegerd , Dagnaes‐Hansen Frederik , Stenram Unne
Published: February 2007

Prostate cancer has a slow growing noninvasive phase, but, in general, is invasive on diagnosis. An initial step in the invasion of surrounding normal tissue is the activity of proteolytic enzymes such as components of the plasminogen activator system (PA). In cell culture, the primary human prostate cancer cell line 1013L expressed no urokinase type‐PA (uPA), while DU 145, a cell line derived from a metastatic lesion, expressed high levels of uPA. The DU 145 cells grew easily as xenografts but the establishment of 1013L in the SCID mice was possible only with the aid of a gelatin sponge (Spongostan). The latency period was 42–64 days, followed by a slow growth phase before a fast growth phase occurred. This fast growth phase was characterized by rapid degeneration of tumor tissue, while high proliferation occurred around the blood vessels. On serial transplantation of tumor material, the growth pattern was similar. Furthermore, the 1013L tumor was encapsulated by connective tissue and no invasiveness could be detected. We found that 1013L tumor homogenates had hardly detectable levels of uPA, i.e., 300‐fold lower than we found in the invasive prostate xenograft DU 145. In addition, no expression of uPA was found in the plasma of 1013L tumor‐bearing mice whilst uPA antigen was detected in the plasma of DU 145 tumor‐bearing mice. In conclusion, the 1013L cell line, which exhibits a nonaggressive pattern, could be a good model for studying progression of prostate cancer to a more aggressive phenotype in vivo and in vitro.
Microvascularity in benign prostatic hyperplasia
Authors: Deering Robert E. , Bigler Steven A. , Brown Marianne , Brawer Michael K.
Published: February 2007

The vascular density of benign prostatic hyperplasia (BPH) has not been characterized. Previously we reported vessel density (vv/mm2) in prostatic carcinoma was twice that of normal prostate [Bigler et al.: Hum Pathol 24:220–226 1993]. To further characterize vessel density in benign prostate tissue we examined 15 cases of BPH obtained by open prostatectomy. Vessels were stained with antibodies to Factor VII‐related antigen, and vessel density was measured using computer‐assisted image analysis.

Vessel density was analyzed between various histologic tissue types. Mean vessel density in all transition zone tissue was 70.2 vv/mm2. Vessel density in epithelial hyperplastic nodules (mean 99.3, SD 40.7) exhibited density levels similar to those found in prostatic carcinoma (mean 101.4, SD 35.6). Vessel density in epithelial nodules was significantly higher than in non‐nodular epithelial tissue (mean 76.7, SD 23.1; P < 0.001, ratio = 1.3). Higher vessel densities were found in hypercellular stromal nodules (mean 64.7, SD 19.1) than in adjacent stromal areas (mean 36.5, SD 15.3; P < 0.001, ratio = 1.8). Overall, vessel density in BPH was higher than previously found in benign tissue measured in radical prostatectomy specimens, especially in areas of nodular morphology.
Synergistic effects of estrogen and androgen on the prostate: Effects of estrogen on androgen‐and estrogen‐receptors, BrdU uptake, immunohistochemical study of AR, and responses to antiandrogens
Authors: Suzuki Kazuhiro , Ito Kazuto , Suzuki Takanori , Yamanaka Hidetoshi , Honma Seijiro
Published: February 2007

To investigate the synergistic effects of estradiol (E2) and testosterone (T) on prostate growth, castrated Wistar rats were treated with 1 mg/day of T or with 1 mg/day of T and 0.01 mg/day of E2 for 6 weeks. The weight of the prostate in T + E2‐treated rats was significantly higher than that in T‐treated and normal rats, parallel with the bromodeoxyuridine (BrdU) labeling index (LI). Nuclear androgen receptor (AR) content in the T + E2 group was significantly higher than that in the T group. But they were lower than that in normal rats. And there were no significant differences between the groups in nuclear estrogen receptor (ER) content. Immunohistochemical studies with the AR antibody revealed positive staining in the prostatic epithelium and stromal cells in the normal, T‐treated, and T + E2‐treated animals. However, castration led to loss of staining.

Response to steroidal antiandrogens was also determined. Antiandrogen treatment abrogated the increases in nuclear AR content and BrdU LI, and prevented immunohistochemical staining.

These results suggest that AR and ER, which were measured in this study, were not indicators of prostatic proliferation. We further need to investigate other factors, including other types of receptors, growth factors involved in epithelial‐stromal interaction, and so on.
Inhibition of prostate cancer growth by estramustine and colchicine
Authors: Fakih Marwan , Replogle Tracy , Lehr Jeff E. , Pienta Kenneth J. , Yagoda Alan
Published: February 2007

Hormone‐refractory prostate cancer continues to be associated with a very poor prognosis. Agents that inhibit microtubule function have been found to be cytotoxic to prostate cancer cells in preclinical and clinical settings. It was the aim of this study to assess the activity of estramustine and colchicine, two microtubule inhibitors, in hormone‐refractory prostate cancer. In clinically achievable concentrations, the combination of estramustine and colchicine was cytotoxic to both the Dunning rat prostate adenocarcinoma cell line MAT‐LyLu (MLL) and human prostate cancer cells (PC‐3). Microtubule function was assessed in vitro to evaluate possible mechanisms of action. In motility and cell cycle analysis assays, estramustine and colchicine inhibited cellular motility but not cell cycle transit. In vivo, these two agents both inhibited the growth of implanted Dunning rat prostate adenocarcinoma MLL cells but did not appear to have additive effects. The use of oral colchicine in the treatment of hormone‐refractory prostate cancer requires further investigation.
Assay variability in serum prostate‐specific antigen determination
Authors: Brawer Michael K. , Daum Phyllis , Petteway Jason C. , Wener Mark H.
Published: February 2007

Consistency and reproducibility of serum prostate‐specific antigen (PSA) measurement are essential in the application of this analyte to early detection or screening programs. In the present investigation, we sought to compare serum PSA levels determined by the IMx assay (Abbott Laboratories, North Chicago, IL) and the Tandem E (Hybritech Inc., San Diego, CA) to determine whether there were differences.

Two hundred twenty‐eight random sera from our archival bank were investigated. One hundred‐eight specimens were in the Tandem range of 2.0–10.0 ng/ml, and prostatic histology was known based on either systematic sector needle biopsy or transurethral resection. PSA was measured with three different lots of the IMx and Tandem assays.

Over the entire range, there was a good correlation (r2 = 0.985); however, in the more useful clinical range of 2.0–10.0 ng/ml, the correlation was reduced to 0.923; in the 2.0–6.0 ng/ml range, it was further reduced to 0.852. The slope for the entire range was 0.948; however, in the 2.0‐ to 10.0‐ng/ml range, it was 0.894; in the 2.0‐ to 6.0‐ng/ml range, the slope was 0.815. Using PSA cutoffs of 4.0, 5.0, and 6.0 ng/ml, significant decrease in abnormal PSA values in men with cancer was observed with the IMx compared with Tandem.

These data suggest that the IMx and the Tandem PSA assays are not equivalent, and in most patients a lower value is realized with the IMx assay. This bias appears to be greater in men with prostate cancer. Clinicians must be aware which assay their patients are being tested with, and laboratory technicians should run internal standards to ensure lack of significant intralot variability and consistency over time. © 1995 Wiley‐Liss, Inc.
Survival of human prostate carcinoma, benign hyperplastic prostate tissues, and IL‐2‐activated lymphocytes in SCID mice
Authors: Lubaroff David M. , Cohen Michael B. , Shultz Leonard D. , Beamer Wesley G.
Published: February 2007

Mice, homozygous for the mutation severe combined immunodeficiency (scid) and also segregating for the mutation hypogonadal (hpg), were tested for their potential use as an in vivo model system for studying the growth of human prostate cancer and benign hyperplastic prostate tissue grafts. Fresh human prostate cancer or benign hyperplastic prostate tissue was implanted subcutaneously into androgen‐replete C.B. 17 scid/scid males, and into androgen‐deficient hpg/hpg scid/scid or androgen‐replete +/? scid scid males. The tissue grafts grew in both androgen‐replete and androgen‐deficient host mice. When dihydrotestosterone (DHT) was administered at tissue grafting, both the incidence and size of the tissue grafts increased. Histology of tissue from tumors in the androgen‐replete hosts showed a well‐differentiated prostate adenocarcinoma, whereas the androgen‐deficient hpg/hpg scid/scid host showed either undifferentiated tumors or adenocarcinomas with few glandular structures. These data suggest the androgen deficient environment selected for growth of androgen‐independent tumor tissue. Finally, when interleukin‐2 (IL‐2)‐activated tumor‐infiltrating lymphocytes were injected into scid/scid hosts, the cells were found to survive and could be identified in the spleen of the recipient mice. These results indicate that growth of human prostate tissues and IL‐2‐activated lymphocytes in scid/scid mice is a viable model system for in vivo studies of prostatic disease. © 1995 Wiley‐Liss, Inc.
PSA assay of dried blood samples from the ear lobe on a filter paper with special reference to prostatic mass screening
Authors: Watanabe Hiroki , Ohe Hiroshi , Saitoh Masahito , Kojima Munekado , Tanaka Toshiyuki , Ito Sachio
Published: February 2007

To eliminate complicated procedures for taking blood samples in the field work for prostatic mass screening, we developed a method for PSA testing using blood samples taken from the ear lobe and dried on a filter paper. The stability, the diagnostic efficacy, and the cost‐benefit effect of the method were confirmed in hospitalized patients, as well as in subjects from our mass screening program. © 1995 Wiley‐Liss, Inc.
Association of some hydrolytic enzymes with the prostasome membrane and their differential responses to detergent and PIPLC treatment
Authors: Fabiani Roberto , Ronquist Gunnar
Published: February 2007

Prostasomes are human prostate derived organelles that were isolated from both prostatic fluid and seminal plasma for the present study. Specific activities were determined for prostasome membrane‐associated enzymes, alkaline phosphatase (ALP), 5′‐nucleotidase (5′NT), and alkaline phosphodiesterase I (APD). The mode of their membranous anchoring was studied by treatment of prostasomes with phosphoinositol‐specific phospholipase C (PIPLC) and different detergents. A substantial amount of ALP (50%) and 5′NT (31%) was released by incubation of prostasomes with 2 U/ml of PIPLC contrary to the small amount of APD (12%) released by the same treatment. After PIPLC treatment, the enzymes were recovered in the aqueous phase after phase repartition in Triton X‐114 indicating that PIPLC removed the hydrophobic domain converting the enzymes from membrane‐linked to aqueous soluble forms. Octyl glycoside was the most efficient one among different detergents to solubilize the enzymes from the prostasome membrane. Both ALP and 5′NT were resistant to the treatment with Triton X‐100 and Triton X‐114. These results suggest that ALP, 5′NT, and APD are more or less extensively linked to the prostasome membrane via a glycophosphoinositide anchor. © 1995 Wiley‐Liss, Inc.
Differentiation‐stage specific expression of oncoprotein 18 in human and rat prostatic adenocarcinoma
Authors: Friedrich Bengt , Grönberg Henrik , Landström Marene , Gullberg Martin , Bergh Anders
Published: February 2007

Oncoprotein 18 (Op18) is an intracellular phosphoprotein that has been shown to be overexpressed in a number of human malignancies. In the present report we have studied the pattern of Op18 expression in normal, hyperplastic, and malignant prostatic tissue as well as in rat prostatic tumor lines. One of the objectives of the present work was to establish whether the level of Op18 expression can be used as a prognostic marker in human prostatic adenocarcinoma. To that end, sections from normal, hyperplastic, and malignant human prostatic tissue were examined by immunohistochemistry for expression of Op18.

In the normal and hyperplastic prostate, Op18 expression was observed in basal glandular epithelial cells, whereas the columnar luminal epithelial cells were not stained by the anti Op18 antibodies. In highly differentiated prostatic cancers occasional epithelial cells were stained, while in poorly differentiated tumors most of the epithelial cells contained Op18 immunoreactivity. The staining pattern was similar in the primary prostatic tumor and in the regional lymph node metastases. Most importantly, a limited survey of prostatic cancer patient samples (n = 40) showed a significant correlation between the fraction of Op18 immunoreactive cells and survival. Studies of a rat prostatic tumor model, showed that only a few cells were stained in the highly differentiated Dunning R3327PAP tumor, while most cells were stained in the anaplastic AT1 rat prostatic tumor. Interestingly, castration of rats resulted in an increased Op18 immunoreactivity, within 14 days, in the highly differentiated rat R3327PAP prostatic tumor.

In conclusion, the level of Op18 expression seems to be related to cellular differentiation, histological grade, and survival in prostatic cancers. These findings show that Op18 immunoreactivity may be useful as a prognostic marker in prostatic cancer. In addition it may help in the differentiation between highly differentiated prostatic tumors and non‐malignant conditions. © 1995 Wiley‐Liss, Inc.
Nuclear morphometry is of independent prognostic value only in T1 prostatic adenocarcinomas
Authors: Vesalainen S. , Lipponen P. , Talja M. , Kasurinen J. , Syrjänen K.
Published: February 2007

A series of 325 patients with prostatic adenocarcinoma followed‐up for over 13 years was subjected to histomorphometric analysis for the following prognostic factors: the Gleason score and 10 nuclear morphometric factors (mean nuclear area, nuclear perimeter, shortest and longest nuclear axis, form factor and their SDs), using the IBAS 1&2 image analyzer. Nuclear factors, Gleason score (P= 0.0013−0.0148), and T‐category (P= 0.004−0.0107) were significantly interrelated, while the M‐category was independent of the morphometric parameters. Nuclear factors (P= 0.0003‐0.5), Gleason score (P< 0.0001), T‐category (P< 0.0001) and M‐category (P< 0.0001) predicted the disease outcome. In T1‐2M0 tumors, the T‐category (P= 0.0001), Gleason score (P< 0.0001), SD of nuclear area (P= 0.057), SD of nuclear perimetry (P= 0.039), mean shortest nuclear axis (P= 0.030), SD of the shortest nuclear axis (P= 0.0045), SD of the longest nuclear axis (P= 0.009), and nuclear form factor (P= 0.0123) were significant prognostic factors. In the multivariate survival analysis, the nuclear area had independent prognostic significance only in T1 tumors, whereas in other subcategories, the clinical stage, Gleason score, and patient age included all the available prognostic information. The results indicate that nuclear morphometric measurements are of borderline significance only in evaluating the intrinsic malignancy of prostatic adenocarcinoma. © 1995 Wiley‐Liss, Inc.
Orthotopic placement of the dunning R3327 AT‐3 prostate tumor in the copenhagen X fischer rat
Authors: Rubenstein Marvin , Saffrin Robert , Shaw Michael , Muchnik Sergey , Guinan Patrick
Published: February 2007

Orthotopic placement of in vitro propagated Dunning R3327 AT‐3 tumor cells resulted in a greater percentage of tumor takes and a two‐fold shift in the exponential growth curve compared to flank implantation. The orthotopic tumor appeared to disseminate preferentially to regional lymph nodes, rather than to the lungs which is characteristic of flank tumors. The results suggest an important role of stromal‐epithelial interactions in the growth of this tumor. © 1995 Wiley‐Liss, Inc.
Diversity and modulation of plasminogen activator activity in human prostate carcinoma cell lines
Authors: Lyon Paul B. , See William A. , Xu Yong , Cohen Michael B.
Published: February 2007

Baseline cellular plasminogen activator (PA) activity, and the cellular proteins responsible for variations in PA activity were evaluated in three human prostate carcinoma cell lines. Net PA activity in the cell lines PC‐3, DU‐145, and LNCaP was measured using a plasminogen‐dependent fibrin lysis assay. These three cell lines were then analyzed to determine the specific protein(s) responsible for differences in PA activity. mRNA and protein levels of cellular urinary PA (uPA), tissue PA (tPA), PA inhibitor 1 (PAI1), PA inhibitor 2 (PAI2), and uPA receptor (uPAr) were measured using Northern analysis and ELISA assays. Net cellular PA activity in the three cell lines varied over a 3‐fold range (PC3 > DU145 > LNCaP). Net PA activity in the fibrinolysis assay demonstrated a direct correlation with mRNA transcript levels of uPA, tPA, PAI1, and uPAr (PC‐3 > DU‐145 > LNCaP). uPA protein was identified in both the PC‐3 and the DU‐145 lines. tPA, PAI1, and PAI2 proteins were identified only in PC‐3 cells. In general, cellular protein levels correlated with mRNA levels. These findings demonstrate that prostate carcinoma cell lines vary in their net PA activity. This variability results from both qualitative and quantitative differences in the cellular expression of PA regulatory proteins. © 1995 Wiley‐Liss, Inc.
Molecular mass and carbohydrate structure of prostate specific antigen: Studies for establishment of an international PSA standard
Authors: Bélanger Alain , Van Halbeek Herman , Graves Howard C. B. , Grandbois Kay , Stamey Thomas A. , Huang Lihua , Poppe Izabella , Labrie Fernand
Published: February 2007

Despite the widely accepted use of prostate specific antigen (PSA) as a marker of prostate cancer, this molecule has not yet been completely characterized. Past studies have well established, however, using both amino acid and cDNA sequencing techniques, that PSA contains 237 amino acids, with a molecular mass of 26,079 Da for the peptide moiety of the molecule. The present study reports analysis of this protein by ion spray mass spectrometry (ISMS) and analysis of its carbohydrate moiety by NMR spectroscopy. The predominant PSA molecular species detected by ISMS was at relative molecular mass (Mr) of 28,430, indicating that PSA contains a carbohydrate residue of Mr 2,351, for a total percentage of carbohydrate of 8.3%. Analysis of PSA by SDS‐PAGE, however, showed a Mr of 32,000 to 33,000, suggesting an overestimation of the molecular weight by the latter technique. The complete primary structure of the PSA carbohydrate chain was determined by NMR spectroscopy in combination with carbohydrate composition analysis. The experimentally determined carbohydrate content of PSA confirms that only one N‐glycosylation site is occupied in the protein. The proposed carbohydrate structure is a diantennary N‐linked oligosaccharide of the N‐acetyllactosamine type, with a sialic acid group at the end of each of the two branches. In addition, our data indicate that ∼70% of the PSA molecules contain a fucose group in the core chitobiose moiety. The calculated molecular weight of this carbohydrate structure (Mr 2,351.8) is in excellent agreement with the predicted molecular weight of the carbohydrate group, based on the Mr 28,430 for PSA measured by ion spray mass spectrometry and Mr 26,079 calculated from the consensus sequence for the peptide portion of the molecule. ISMS of PSA is thus proposed as a convenient and reliable method of quality control, an indispensible step towards international standardization of this very important tumor marker for detection and monitoring of prostatic diseases, especially prostate cancer. © 1995 Wiley‐Liss, Inc.
Identity of PSA purified from seminal fluid by different methods: Comparison by amino acid analysis and assigned extinction coefficients
Authors: Stamey T. A. , Teplow D. B. , Graves H. C. B.
Published: February 2007

To determine the true extinction coefficient of prostate specific antigen (PSA) and to measure any differences in PSA when isolated from seminal fluid by four different published methods, we studied 10 different lots of PSA by quantitative amino acid analysis.

Despite an expected PSA concentration of 1 mg/ml based on gravimetric analysis at an average optical density of 1.45 at 280 nm, we recovered only 0.79 mg/ml by quantitative amino acid analysis (range 0.752 to 0.820 mg/ml with a coefficient of variation [C.V.] of 3.3% among the 10 lots). The concentration of 0.79 mg/ml was based on a molecular weight of 28,430 daltons for glycosylated PSA determined by ion spray mass spectroscopy [Bélanger et al: Prostate, 27:187–197, 1995]. From these 10 amino acid analyses, we calculated the extinction coefficient of PSA at 280 nm as 1.84 ± 0.04 ml × mg−1 × cm−1 (range 1.78 to 1.90 with a C.V. of 2.2%). © 1993 Wiley‐ Liss, Inc.

Similar concentrations of PSA were obtained by amino acid analysis regardless of the method of purification. These observations support the presence of a single from of PSA in seminal fluid and are consistent with the molecular evidence that PSA is transcribed from a single gene locus on the long arm of chromosome 19 [Riegman et al.: Genomics 14:6‐11, 1992]. They do not support the recent contention by the Roswell Park group that the PSA they isolated in 1979 [Wang et al.: Invest Urol 17:159‐163, 1979; Wang et al.: Prostate 24:107‐108, 1994] is different from p30 reported a year earlier [Sensabaugh: J Forensic Sci 23:106–115, 1978]. © 1993 Wiley‐ Liss, Inc.