Changes of Urinary Nerve Growth Factor and Transforming Growth Factor beta-1 in Male Patients with Lower Urinary Tract Symptom / 대한비뇨기과학회지

PURPOSE: Increased expression of the nerve growth factor (NGF) in the urinary bladder may contribute to irritative bladder symptoms in patients with benign prostatic hyperplasia. It has been reported that the transforming growth factor beta (TGF-beta) can initiate NGF synthesis. This study investigated the changes in the urinary NGF and TGF-beta1 in male patients with lower urinary tract symptoms. MATERIALS AND METHODS: The study groups included 25 male patients with lower urinary tract symptom and 10 normal male patients used as the reference controls. The evaluation included history taking, urinalysis, international prostatic symptom score (IPSS), and urodynamics. Voided urine was collected in all patients. The urinary NGF and TGF-beta1 concentration was analyzed by using an enzyme linked immunosorbent assay and these results were compared with the control group. RESULTS: From the analysis of the IPSS, the obstructive and irritative symptom scores were higher. The urodynamic study showed that more than half of the patients had detrusor instability and bladder outlet obstruction (BOO). The NGF and TGF-beta1 urinary concentration were significantly higher in patients with lower urinary tract symptoms compared to the control. Furthermore, the NGF and TGF-beta1 urinary concentration were significantly higher in patients with BOO compared with patients with non-BOO. CONCLUSIONS: NGF and TGF-beta1 may play a role in the lower urinary tract symptoms and these changes can be detected in urine samples. Therefore, these growth factors may be used as markers to evaluate lower urinary tract symptoms.

Effects of Epidermal Growth Factor, Basic Fibroblast Growth Factor and Keratinocyte Growth Factor on PSA Secretion and PSA mRNA Expression in LNCaP Prostate Cancer Cell Line / 대한비뇨기과학회지

PURPOSE: In order to characterize the effects of growth factors (EGF, bFGF, KGF) on the regulation of the PSA secretion and the PSA mRNA expression of androgen- dependent LNCaP prostate cancer cell line in serum-free conditions. MATERIALS AND METHODS: LNCaP cells at a concentration of 1x104cells/well, suspended in T-medium containing 2% TCM, were seeded in 24 well plates and were exposed to four different concentrations of these growth factors to evaluate the molecular basis of PSA secretion. Cell numbers were evaluated by crystal violet assay on day 5. PSA concentrations in conditioned medium were determined on day 5, and PSA/cell number was also calculated to measure net PSA secretion per cell. PSA mRNA expression of LNCaP was assessed by RT-PCR and Northern blot analysis on day 5. RESULTS: bFGF and KGF had significant stimulatory effects (p<0.05) on the proliferation of LNCaP. However, EGF had minimal, not significant, growth stimulatory effects. EGF, bFGF and KGF did not increase the PSA secretion of LNCaP and no apparent increase or decrease in the steady-state levels of the PSA mRNA expression of LNCaP could not be detected in spite of addition of EGF, bFGF and KGF. CONCLUSIONS: bFGF and KGF, not EGF, directly stimulate the proliferation of LNCaP cells. However, bFGF and KGF as well as EGF do not affect the PSA secretion and the PSA mRNA expression of androgen-dependent LNCaP in the absence of androgenic milieu. The regulation of the PSA secretion and the PSA mRNA expression of LNCaP is not directly associated with EGF, bFGF and KGF.

Effects of Frequency and Urgency on Urinary Nerve Growth Factor and Transforming Growth Factor beta-1

PURPOSE: Frequency and urgency are common symptoms in women. Growth factors have been reported to play a role in these irritative symptoms. This study was performed to investigate the changes in urinary growth factors according to frequency and urgency in adult female patients. MATERIALS AND METHODS: The study groups included 30 patients with frequency and urgency and 10 normal patients as reference controls. Evaluation included history taking, urinalysis, voiding diary, and urodynamics. Voided urine was collected. The urinary concentration of nerve growth factor(NGF) and transforming growth factor beta-1(TGF-beta1) were analyzed and these results were compared with control groups. RESULTS: From the analysis of voiding diary, the degree of frequency was increased and functional bladder capacity was decreased in the frequency and urgency patients. Also, capacity for first voiding sense and full capacity were decreased in frequency and urgency patients from urodynamic study. The urinary concentration of NGF and TGF-beta1 were significantly increased in frequency and urgency patients compared with control patients. CONCLUSIONS: NGF and TGF-beta1 may play a role in irritative symptoms such as frequency and urgency and these changes can be detected in urine sample. Thus, these growth factors may be a marker to evaluate bladder function.

Epidermoid Cyst of Testis / 대한비뇨기과학회지

A 24-year-old man visited our hospital with painless scrotal mass 2 weeks in duration. Physical examination revealed about 3cm in diameter, ovoid, hard, movable and painless mass in the left side of the scrotum. He had no history of trauma, voiding difficulty or infection sign. CBC, blood chemistry and tumor marker were normal. Left orchiectomy was done. Tumor was a well encapsulated white-gray ovoid mass filled with yellowish gray material and histogically diagnosed as epidermoid cyst. Benign tumors of testis are rare, acounting for less than 1% of all testicular tumors. Herein we report a case of epidermoid cyst of testis and review the literatures.

Permanent Genotypic and Phenotypic Change of Prostate Cancer Cell Line LNCaP through Cellular Interactions with Prostate or Bone Fibroblasts in vitro or in vivo / 대한암학회지

PURPOSE: Cell-cell interactions determine normal prostate development and subsequent neoplastic transfor mation. The progression of prostate cancer from androgen-dependent to androgen-independent states involves multiple steps of genetic changes mediated by tumor-microenvironment interactions. To understand the epigenetic factors that lead to progression, we studied if 1) androgen-dependent and non-metastatic LNCaP may interact with prostate or bone fibroblasts under microgravity-simulated conditions in vitro. 2) LNCaP may interact with prostate fibroblasts in vivo, and acquire androgen-independence and metastatic potential. MATERIALS AND METHODS: The LNCaP sublines were generated as follows. 1) LNCaP cells were grown in vitro either alone or with prostate or bone fibroblasts under microgravity-simulated conditions. 2) LNCaP cells were grown in vivo as chimeric tumors with prostate fibroblasts. The LNCaP sublines were characterized by studies of chromosomal analysis, comparative genomic hybridization and, in vivo tumorigenicity and metastatic potential. RESULTS: In comparison to the parental LNCaP cells, the LNCaP sublines underwent permanent genotypic and phenotypic changes manifested in androgen-independence and metastatic potential. CONCLUSION: These results emphasize the importance of cell-cell interaction as a critical determinant that could "induce" or "select" progenies favoring enhanced prostate cancer growth and progression. This concept favors the development of toxic gene therapy targeting both prostate cancer epithelium and supporting bone stroma for an effective eradication and prevention of prostate cancer bone metastasis.

3-Dimensional Culture of Prostate Organoid under Microgravity-Simulated Growth Conditions: Cell-Cell Interactions Favoring Cancer Growth and Progression / 대한비뇨기과학회지

PURPOSE: Three-dimensional(3-D) organization is critical for both the normal development and, tumor growth and progression. Cell-cell and cell-matrix interactions determine normal prostate development and its subsequent neoplastic transformation. To understand the epigenetic factors that lead to cell transformation, a 3-D human prostate cell culture was established with prostate epithelial cells grown in a rotating-wall vessel(RWV) under microgravity-simulated conditions with either alone or with prostate or bone stromal cells. We tested the hypothesis of whether phenotypic and genotypic alterations of LNCaP cells may be achived when grown as 3-D organoids. MATERIALS AND METHODS: LNCaP cells were seeded in RWV alone and with either human prostate or bone fibroblasts. After period of 2 months, RWV1, 2, and 3 cell lines were established from the prostate organoids and were characterized. RESULTS: While LNCaP cells injected orthotopically failed to form tumors in castrated mouse, RWV-derived cell lines formed PSA-producing tumors and metastasized to lymph node, bone, lung and liver, which stained positively by PSA antibody. RWV cells grew faster than parental LNCaP, especially in sex hormone-free conditions. Unlike parental LNCaP cells which respond positively to androgen and estrogen-induced growth and PSA expression, RWV cells are insensitive to sex steroid-induced growth, but remain sensitive to androgen for induction of PSA expression. Comparative genomic hybridization(CGH) results demonstrated that RWV cell lines have different chromosomal gain and loss each other as compared to those of LNCaP. CONCLUSIONS: Cell-cell and cell-matrix interactions of androgen-dependent and non-metastatic LNCaP cultured alone or with either prostate or bone fibroblasts in 3-D culture under microgravity-simulated conditions resulted in induction of androgen- independent and metastatic LNCaP sublines, RWV cell lines, meaning androgen- independent progression. Phenotype and genotype of RWV cell lines are definitely dissimilar to those of parental LNCaP. 3-D culture of prostatic epithelial cells under microgravity-simulated conditions could be novel approach to the study of normal development and cancer of prostate as an ideal in vitro model and, will be further exploited.

Effects of Lipopolysaccharide on Inflammation within the Bladder Muscle and Muscle Contractility in the Rats / 대한비뇨기과학회지

PURPOSE: The mechanisms, which alter urinary bladder muscle function during infectious cystitis, are poorly understood. The objective of this study was to identify potential resident targets for endotoxin lipopolysaccharide(LPS) within normal bladder smooth muscle and test the hypothesis that LPS induces an inflammatory response within the bladder muscle and that this is associated with a decrease in muscle contractility. MATERIALS AND METHODS: Male rats were studied 24 hours after a single bolus i.p. injection of LPS(15mg/kg). Whole-mount preparations of the bladder muscle were immunohistochemically stained for neutrophils(myeloperoxidase), macrophages(ED2), activated leukocytes(LFA-1) and mast cells(FITC-Avidin). Contractile activity was assessed from muscle strips of the bladder in response to bethanechol(0.3-300microM). Voiding frequency and urine volume for 24 hours were measured using metabolic cage. Cystometry was performed to measure the intravesical bladder pressure. RESULTS: Using the resident macrophage marker ED2, dense network of macrophages were observed within the bladder muscle of control and LPS treated rats. Few neutrophils(myeloperoxidase-positive cells, 2.3 +/- 0.38 cells, x200) were detected in whole-mounts of bladder muscle of control rats, while LPS pretreatment resulted in a significant increase in the number of neutrophils which demonstrate inflammatory response(10.8 +/- 1.70 cells, x200, p<0.001). LFA-1 immunohistochemistry demonstrated an increased presence of LFA-1 positive cells in bladder muscle of LPS treated rats, which had a morphology similar to both neutrophils and resident macrophages. The expression of LFA-1 is known as a marker of cells that are in an activated state. LPS pretreatment resulted in a significant reduction in bladder muscle contractions in response to bethanechol(i.e. control = 0.049 +/- 0.010 vs. LPS= 0.029 +/- 0.003 gr/mm2/sec, 100microM, p<0.05). Voiding frequency of LPS treated rats was significantly increased compared to that of control rats. In LPS treated rats, voiding phase representing bladder contractility in cystometry was observed. CONCLUSIONS: These data demonstrate that a single intraperitoneal injection of LPS initiates an inflammatory response within the bladder muscle that is associated with a decrease in the functional activity of the bladder. We hypothesize that secretions from the resident macrophages and extravasated leukocytes within the muscle cause the observed suppression in bladder muscle activity in vitro.

Derivation of Androgen-Independent LNCaP Sublines from Cell-Cell Interactions of LNCaP and Human Prostate Fibroblasts in vivo / 대한비뇨기과학회지

PURPOSE: A cell-cell interaction in which in vivo inoculation of androgen-dependent, non-tumorigenic LNCaP and human bone fibroblast resulted in derivation of androgen-independent and metastatic LNCaP subline(C4-2) in castrated hosts. The purpose of this study is to evaluate if human prostate fibroblasts when grown together with LNCaP may promote androgen-independent growth and enhance metastatic potential. MATERIALS AND METHODS: LNCaP cells and human prostate fibroblasts derived either from peripheral or transition zone co-inoculated in athymic mice for 8 weeks, and then mice were castrated. The chimeric tumors were maintained for additional 4 weeks. The LNCaP sublines, designated P4 and T4, were established and characterized. These sublines were co-inoculated again in castrated mice with human prostate fibroblasts for 8-12 weeks. And then second generation LNCaP sublines, P4-2 and T4-2, were established and also characterized. RESULTS: Marked cytogenetic alterations were observed in P4-2, P4, T4-2 and T4 LNCaP sublines in comparison to parental LNCaP. Although LNCaP cells injected orthotopically did not form tumors in castrated hosts, LNCaP sublines formed PSA-producing tumors and had metastatic potentials to lymph node, lung, liver and bone. These P and T sublines had androgen-independent growth characteristics and metastatic potential. CONCLUSIONS: Cell-cell interactions between prostatic epithelium and their surrounding fibroblasts could contribute to androgen-independent characteristics and enhanced metastatic potential of localized prostate cancer in vivo.

Effects of Various Growth Factors on the Growth and PSA Secretion of the Androgen-dependent LNCaP Prostate Cancer Cell Line / 대한비뇨기과학회지

PURPOSE: The growth and PSA secretion of prostate carcinoma is predominantly regulated by androgens. However, locally produced growth factors(GFs) also have been shown to play a crucial role in the proliferation of androgen-dependent prostatic tumor cells. Androgen has been proposed to stimulate the cell proliferation and PSA secretion by modulating the activity of some of these growth factors. The objectives of this study are to determine the effects of various GFs (epidermal growth factor; EGF, basic fibroblast growth factor; bFGF, keratinocyte growth factor; KGF, hepatocyte growth factor; HGF) on the growth and PSA secretion of androgen-dependent LNCaP prostate cancer cell line. MATERIALS AND METHODS: To determine the effects of EGF, bFGF, KGF and HGF on the growth and PSA secretion of LNCaP, LNCaP cells at a concentration of 3 x 103 cells/well, suspended in T-medium containing 2% TCM, were seeded in 96 well plates. Cells were exposed to six different concentrations (0.5, 1, 5, 10, 20 and 40 ng/ml) of GFs. Cell numbers were evaluated by crystal violet assay on day 3, 5 and 7, and PSA concentrations in conditioned medium were determined on day 5. RESULTS: EGF and HGF had minimal, not significant, stimulatory effects on the growth of LNCaP. However, bFGF and KGF had significant growth stimulatory effects (P<0.05). EGF, bFGF, KGF and HGF did not have any stimulatory effects on the PSA secretion of androgen-dependent LNCaP cell line. CONCLUSIONS: bFGF and KGF, not EGF and HGF, directly stimulate the growth of LNCaP cells. However, bFGF and KGF as well as EGF and HGF do not affect the PSA secretion of LNCaP. There seems to be another signal transduction pathway, which is not associated with GFs mentioned above, for PSA secretion of androgen-dependent LNCaP cell line.

The Effect of A Potent Calcium Channel Blocker, Nifedipine, on the Castration-induced Apoptosis of the Rat Ventral Prostate / 대한비뇨기과학회지

The rapid involution of the rat ventral prostate after castration is an active process initiated by removal of the inhibitory effects of androgen on prostatic cell death. The degradation of genomic DNA into nucleosomal-sized fragments if an early event in this process and is catalyzed by Ca2+Mg2+-dependent endonuclease activity which is dependent upon calcium ions. The morphologic correlation of the involution process involves a series of structural changes which are collectively referred to as apoptosis. Since the castration-induced endonuclease is dependent upon calcium ions for maximal activity, a potential involvement of a intracellular calcium in the castration-induced prostatic cell death was investigated. Acute disturbance in intracellular calcium homeostasis within the ventral prostate by means of a potent calcium channel blocker, nifedipine, simultaneous with castration resulted in a significant decrease in prostatic apoptosis. This result points to a potential role intracellular calcium levels in the mechanism of activation of castration-induced death of the androgen-dependent epithelial cells in the ventral prostate.

Wegener`s Granulomatosis of the Ureter Mimicking Ureteral Tumor / 대한비뇨기과학회지

Wegener`s granulomatosis (WG) is a multisystem disorder of unknown origin characterized by necrotizing granulomatous vasculitis. WG usually affects the upper respiratory tract, lungs, and kidneys with focal glomerulonephritis, but almost any organ can be affected. The ureter is primarily an unusual location for lesions of WG. A 30-year-old woman presented with intermittent right flank pain and hematuria. A renal ultrasound demonstrated unilateral hydronephrosis and a retrograde pyelography revealed a filling defect at right mid ureter and a computed tomography displayed marked concentric thickening of the right ureteral wall which was mimicking ureteral tumor. At nephroureterectomy, the right ureter was found to be obstructed by dense, intramural fibroinflammatory reaction. There was a necrotizing granulomatous vasculitis in the muscle layer of the ureter. Our case represents the rare occurrence of WG presenting ureteral bstruction.

Modification of Epidermal Growth Factor Receptor (EGFR) Expression after Aminoglycoside-induced Acute Tubular Necrosis in Rat / 대한비뇨기과학회지

Acute tubular necrosis induced by aminoglycoside-antibiotics is followed by a regenerative Process which leads to the restoration of damaged tubules. It is well known that tubular regeneration is mediated by polypeptide growth factors such as epidermal growth factor (EGF) In the absence of nephrotoxic alterations, EGF is immunolocalized in distal tubules, whereas epidermal growth factor receptor(EGFR) immunostaining is localized in proximal tubules. After acute tubular necrosis, the sign of regeneration is accompanied by redistribution of EGF immunoreactivity from distal to proximal tubules and a reduction of total immunoreactive EGF due to a decrease of tissue-bound proEGF. However, it is controversial whether EGFR is down- or up-regulated during this regenerative process. We evaluated the time course of the regenerative response subsequent to tubular damage induced by aminoglycoside, with a particular attention paid to EGFR in order to examine whether it is down- or up-regulated. Female Sprague-Dawley rats were treated during 4 and 8 consecutive days with a daily dose of 80 mg/kg gentamicin i.p. Groups of experimental animals (n=10) were terminated at increasing time intervals (5, 9, 12, 16 days) after cessation of treatment. One hour before sacrifice, each individual received i.p. 200mg/kg Blood for the immunohistochemical demonstration of cell proliferation (S-phase cell). Blood was collected at the time of sacrifice to measure serum creatinine and BUN levels. EGFR immunoreactivity was revealed on paraffin-embedded tissue section by immunohistochemical staining using polyclonal anti-EGFR antibody. Upon immunostaining sections in control kidneys, immunoreactive EGFRs exhibited a quite specific and restricted distribution since they were confined to proximal tubules. But proximal tubules undergoing regenerative repair were characterized by a disappearance of EGFR, which expressed BrdU in immunohistochemical sections for BrdU. Beyond the episode of tubular regeneration, proximal tubules recovered immunoreactive EGFR. The results suggest that the apparent loss of EGFR could be due to a process of receptor down-regulation in proximal tubules displaying evidence of regenerative response.

A Clinical Observation of 13 Testicular Embryonal Carcinomas / 대한비뇨기과학회지

We reviewed 13 patients with testicular embryonal carcinoma from July 1982 to May 1994. Embryonal carcinoma accounted for about 25% of total testicular tumors(13/56) and about 34% of nonseminomatous germ cell tumors(13/38). Among the patients with embryonal carcinoma, about 85% were diagnosed in the 15-to-34 year age group. About seventy percent of the patients had metastatic disease at the time of diagnosis and 66.7% of these had distant metastasis including by lung, bone, retroperitoneal lymph node and cervical lymph node, attesting to the aggressiveness of embryonal carcinoma and its tendency to early hematogenous spread. Serum AFP was elevated in 10 patients(76.9%) and hCG in 6 patients(46.2%). All patients with stage I (4/13) were treated with radical orchiectomy only, and all patients with stage II(3/13) were treated with radical orchiectomy and retroperitoneal lymph node dissection(RPLND) followed by chemotherapy. Of the patients with stage III(6/13), 4 patients were treated with radical orchiectomy and chemotherapy, and 2 patients with radical orchiectomy and early chemotherapy, followed by RPLND. The pathologic findings of lymph node at the time of RPLND in stage III were 1 residual embryonal carcinoma and 1 fibrosis. In stage I and II, all patients were still alive. In stage III, 2 patients were still alive for 22 and 48 months. Among the 4 expired patients, 2 were from lung metastasis and the others from sepsis might caused by chemotherapy. These results suggest that the radical orchiectomy only with close clinical observation for stage l had a good survival, and long term survival in stage II patients treated with radical orchiectomy and RPLND followed by chemotherapy will be expected. And in stage III, if the side effects of the chemotherapy is reduced, the better survival may be obtained.

Antisperm Antibody Detection by Immunobead Test (IBT) after Vasectomy Reversal: A Preliminary Study / 대한비뇨기과학회지

A group of 32 men, who had undergone vasectomy reversal and had been infertile for at least 1 year in spite of normal semen characteristics, were tested with direct immunobead test ( IBT) for immunoglobulin (Ig) A and IgG antisperm antibodies bound to sperm surface. A test was positive if 50 % of the motile sperm had 2 attached beads. The overall positive rate of antisperm antibodies was 25% (8/32) and the positive rate of direct test for IgA and IgG antisperm antibodies was as follows: for IgA 12.5% ( 4/32) and IgG 25% (8/32). Of the 8 IBT positive patients, IgG antisperm antibodies were present in all patients with positive direct IBT. So it was suggested that presence of localized IgG antisperm antibodies made a important role in infertility after vasectomy reversal. The majority of IgA and IgG binding was directed to the head and entire sperm surface with a minor incidence to the tail. Clinical definition of direct IBT results and exact role of antisperm antibodies need to be developed in conjunction with future fertility rates of IBT positive patients.

A Clinical Observation on Urothelial Tumor of the Upper Urinary Tract / 대한비뇨기과학회지

urothelial tumors of the upper urinary tract are relatively uncommon, mostly malignant. 25 patients with urothelial tumor of upper urinary tract seen at St. Mary`s hospital from Jan. 1975 to Dec. 1986 are reviewed retrospectively. The average age was 57.3 years, approximately half of them being more than 60 years old. Gross Hematuria was the most prevalent sign and cytologic examination of the urine has proved not to be diagnostic. The most common findings of excretory urography and retrograde urography were non- visualization of the kidney and filling defect. CT can help to evaluate the extent of disease by determining the invasion or metastasis to surrounding tissues or retroperitoneal node. All 25 patients were surgically explored, 23 patients underwent nephroureterectomy and 6 of them was also done lymphadenectomy, distal ureterectomy with ureteral reimplantation was done in 1 patient and simple nephrectomy was done under the misdiagnosis of renal tuberculosis in 1 patient. There were close relations between histological grade and pathologic grade. Of the 25 patients, stage A and grade II were the most common, each of them were 11 and 16 cases. The over-all 2 year survival rate of 13 patients followed up was seventy percent.