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Supplementary Figure 1 from Molecular Features of Hormone-Refractory Prostate Cancer Cells by Genome-Wide Gene Expression Profiles

Supplementary Table 1 from Molecular Features of Hormone-Refractory Prostate Cancer Cells by Genome-Wide Gene Expression Profiles

Supplementary Figure 1 Legend from Molecular Features of Hormone-Refractory Prostate Cancer Cells by Genome-Wide Gene Expression Profiles

One of the most critical issues in prostate cancer clinic is emerging hormone-refractory prostate cancers (HRPCs) and their management. Prostate cancer is usually androgen dependent and responds well to androgen ablation therapy. However, at a certain stage, they eventually acquire androgen-independent and more aggressive phenotype and show poor response to any anticancer therapies. To characterize the molecular features of clinical HRPCs, we analyzed gene expression profiles of 25 clinical HRPCs and 10 hormone-sensitive prostate cancers (HSPCs) by genome-wide cDNA microarrays combining with laser microbeam microdissection. An unsupervised hierarchical clustering analysis clearly distinguished expression patterns of HRPC cells from those of HSPC cells. In addition, primary and metastatic HRPCs from three patients were closely clustered regardless of metastatic organs. A supervised analysis and permutation test identified 36 up-regulated genes and 70 down-regulated genes in HRPCs compared with HSPCs (average fold difference > 1.5; P < 0.0001). We observed overexpression of AR, ANLN, and SNRPE and down-regulation of NR4A1, CYP27A1, and HLA-A antigen in HRPC progression. AR overexpression is likely to play a central role of hormone-refractory phenotype, and other genes we identified were considered to be related to more aggressive phenotype of clinical HRPCs, and in fact, knockdown of these overexpressing genes by small interfering RNA resulted in drastic attenuation of prostate cancer cell viability. Our microarray analysis of HRPC cells should provide useful information to understand the molecular mechanism of HRPC progression and to identify molecular targets for development of HRPC treatment. [Cancer Res 2007;67(11):5117–25]

Aim: Docetaxel-based chemotherapy against castration-resistant prostate cancer (CRPC) has recently been shown to be effective and tolerable. The objective of this study was to retrospectively evaluate the efficacy and toxicity of low-dose docetaxel in combination with dexamethasone. Methods: Thirty-seven CRPC patients were administered a treatment regimen consisting of 50 mg/m2 docetaxel once every 3-4 weeks and 1 mg dexamethasone daily at our institution, between November 2004 and April 2014. Results: Twenty-four patients (65%) had a decrease in serum prostate-specific antigen (PSA) >50%. The median overall survival (OS) and PSA progression-free survival were 26.2 and 10.0 months, respectively. Ten of 12 patients (83%) taking analgesic agents reduced their intake because of decreased pain levels. Grade 3 febrile neutropenia occurred in 2 patients (5%). Nonhematological toxicities were less frequent but sometimes severe. Treatment-related death occurred in 2 octogenarian patients, 1 due to gastric bleeding and the other due to infective endocarditis. Conclusion: Low-dose docetaxel in combination with dexamethasone is feasible in Japanese CRPC patients. Hematological toxicity is less than that seen with standard docetaxel therapy, but it is necessary to monitor patients for severe nonhematological toxicities, particularly very elderly patients.

Combined gemcitabine and cisplatin (GC) treatment is a first line chemotherapy for bladder cancer. However, acquired resistance to GC has been a major problem. To address the mechanism of gemcitabine resistance, and to identify potential biomarkers or target proteins for its therapy, we aimed to identify candidate proteins associated with gemcitabine resistance using proteomic analysis. We established gemcitabine-resistant human bladder cancer cell lines (UMUC3GR and HT1376GR) from gemcitabine-sensitive human bladder cancer cell lines (UMUC3 and HT1376). We compared the protein expression of parental and gemcitabine-resistant cell lines using isobaric tags for relative and absolute quantification (iTRAQ) and liquid chromatography tandem mass spectrometry. Among the identified proteins, ethylmalonyl-CoA decarboxylase (ECHDC1) expression was significantly increased in both of the gemcitabine-resistant cell lines compared to the respective parental cell lines. Silencing of ECHDC1 reduced ECHDC1 expression and significantly inhibited the proliferation of UMUC3GR cells. Furthermore, silencing of ECHDC1 induced upregulation of p27, which is critical for cell cycle arrest in the G1 phase, and induced G1 arrest. In conclusion, ECHDC1 expression is increased in gemcitabine-resistant bladder cancer cells, and is involved in their cell growth. ECHDC1, which is a metabolite proofreading enzyme, may be a novel potential target for gemcitabine-resistant bladder cancer therapy.

Cisplatin is currently the most effective anti-tumor agent available against bladder cancer. To clarify the mechanism underlying cisplatin resistance in bladder cancer, the present study examined the role of the aldo-keto reductase family 1 member C2 (AKR1C2) protein on chemoresistance using a human bladder cancer cell line. The function of AKR1C2 in chemoresistance was studied using the human HT1376 bladder cancer cell line and the cisplatin-resistant HT1376-CisR subline. AKR1C2 was expressed in HT1376-CisR cells, but not in the parental cells. The effect of small interfering (si) RNAs and an inhibitor targeting AKR1C2 was examined to determine whether cisplatin sensitivity can be rescued by blocking AKR1C2 expression or function. Silencing of AKR1C2 mRNA or inhibition of AKR1C2 by 5β-cholanic acid resulted in a decrease in the survival of cells following cisplatin exposure. Intracellular accumulation of reactive oxygen species (ROS) was determined using a 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) fluorescent probe. Cisplatin exposure increased the level of intracellular ROS in HT1376 cells in a dose-dependent manner. The ROS levels in HT1376-CisR cells were significantly lower than those in HT1376 cells and knockdown of AKR1C2 mRNA significantly restored ROS levels. Cisplatin exposure did not increase intracellular ROS in HT1376-CisR cells, although the level of intracellular ROS increased in HT1376 cells following cisplatin exposure. Silencing of AKR1C2 mRNA restored the ROS increase response to cisplatin and menadione as an oxidative stressor in HT1376-CisR cells. Menadione has the function of an oxidative stressor. The silencing of AKR1C2 mRNA restored the increased ROS response to cisplatin and menadione in HT1376-CisR cells. These results indicate that induction of AKR1C2 in human bladder cancer cells aids in the development of cisplatin resistance through antioxidative effects. The results of this study indicate that AKR1C2 may be an effective molecular target for restoring cisplatin resistance.

Background Leiomyosarcomas typically originate in smooth muscle cell. Leiomyosarcoma potentially arising from the adrenal gland is an extremely rare mesenchymal tumors associated with delayed diagnosis and poor prognosis. Case presentation A 34-year-old man visited our department complaining of right hypochondriac pain. Computed tomography demonstrated a solid mass measuring 5.2 cm in diameter above the right kidney, corresponding to the right adrenal gland, and a lymph node mass, which appeared to have invaded the IVC wall. Right adrenalectomy and lymphadenectomy were performed. A microscopic examination revealed primary adrenal leiomyosarcoma with lymph node metastasis. No adjuvant therapy was performed, and the patient remains recurrence-free at 10 months postoperatively. Conclusions We experienced a very rare case of primary adrenal leiomyosarcoma. Aggressive surgical resection including vascular reconstruction may be associated with improved survival.

Bladder cancer incidence increases with age, presumably reflecting a cumulative exposure to carcinogens and ever-increasing life expectancy. While aberrant protein expression due to DNA mutations is an essential step during oncogenesis, one recent interest has been the role of epigenetic changes in regulating bladder tumor development. Because aberrant histone acetylation has been linked to malignant diseases in several cases, histone deacetylase inhibitors have great potential as new anticancer drugs owing to their ability to modulate transcription and induce differentiation and apoptosis. We herein review the current knowledge on epigenetic issues in bladder cancer, particularly regarding histone acetylation, and discuss its implications for understanding the molecular basis and treatment of this disease.

PURPOSE: Efficacy and tolerability of docetaxel-based chemotherapy against hormone-refractory prostate cancer (HRPC) has been shown lately. The objective of this study was to evaluate retrospectively the efficacy and toxicity of low-dose docetaxel in combination with dexamethasone. PATIENTS AND METHODS: Sixteen patients, with a median age of 69.5 years (range 54-85 years), diagnosed with HRPC were administered a treatment regimen consisting of docetaxel (60-80 mg/body or 50 mg/m2) once every 3 or 4 weeks and dexamethasone 1 mg daily at our institution between November, 2004 and March, 2010. RESULTS: The patients received a median of 11.5 cycles of treatment (range, 2-35 cycles). Eleven of 16 patients (68.8%) had a > or = 50% decrease in serum prostate-specific antigen. The median progression-free survival and overall survival times were 7.1 and 20.3 months, respectively. Grade 3 neutropenia occurred only in 2 patients. Infective endocarditis, gastrointestinal or cerebral hemorrhage, and compressive fracture were occurred in each patient. CONCLUSIONS: The combination of low-dose docetaxel every 3-4 weeks and dexamethasone daily was effective and well tolerated in patients with HRPC. However, it is necessary to pay continuous attention to side effects due to the frequent presence of comorbid diseases particularly in the elderly.

We report a rare case of polyostotic fibrous dysplasia on endocrine hyperfunction with elevated human growth hormone and normal serum level of prolactin. There were some differential points of gender, gigantism, endocrine function, and GNAS gene from McCune-Albright syndrome. Malignant transformation was suspected in the pelvic tumor from imaging because rapid growth of the tumor by imaging was observed; however, no malignant change occurred in this case.

A 29-year-old man was admitted to our hospital due to bilateral testicular tumors. The bilateral testicular tumors were palpated and visualized by ultrasound and magnetic resonance imaging (right 8 cm, left 6 cm in diameter). Bilateral high orchiectomies were performed after frozen storage of the sperm. Pathological examination revealed seminoma and immature teratoma in the right testis and seminoma in the left testis. Three months later, computed tomography (CT) showed multiple metastatic lung nodules. In addition, positron emission tomography (PET)-CT examination revealed peri-caval lymph node metastasis together with the lung metastases. Lung metastases disappeared, but the lymph node metastasis reduced and remained after 3 courses of combination chemotherapy with bleomycin, etoposide and cisplatin. PET-CT examination revealed that no uptake of FDG was seen in the lung and lymph nodes. Retroperitoneal lymph node dissection was performed. No viable tumor cells were histologically present in the excited lymph nodes. The postoperative course was good and uneventful at 10 months under androgen replacement therapy without disease recurrences.

Early phase prostate cancer is usually androgen-dependent, with the androgen/androgen receptor (AR) signaling pathway playing a central role. At this stage, the cancer responds well to androgen ablation therapy, but prostate cancers eventually acquire androgen independence and more aggressive phenotypes. Several studies, however, have shown that the majority of tumors still express functional AR, which is often amplified and mutated. To determine if the AR is a plausible therapeutic target, we investigated the anti-tumor effect of small interfering RNAs targeting the AR (siAR) in the human prostate cancer cells, LNCaP and 22Rv1, which express mutated AR. In both types of cells, transfection of siAR suppressed mutated AR expression and significantly reduced cell growth. Furthermore, atelocollagen-mediated systemic siAR administration markedly inhibited the growth of 22Rv1 cells subcutaneously xenografted in castrated nude mice. These results suggest that the AR is still a key therapeutic target even in androgen-independent prostate cancer (AIPC). Silencing of AR expression in AIPC opens promising therapeutic perspectives.

Benign prostatic hyperplasia (BPH) is an age-related disease of unknown etiology characterized by prostatic enlargement and coinciding with distinctive alterations in tissue histomorphology. To identify the molecular mechanisms underlying the development of BPH, we conducted a DNA microarray study using a previously described animal model in which chronic α(1)-adrenergic stimulation by repeated administration of phenylephrine evokes histomorphological changes in the rat prostate that resemble human BPH. Bioinformatic tools were applied to microarray data obtained from prostate tissue to construct a network model of potentially relevant signal transduction pathways. Significant involvement of inflammatory pathways was demonstrable, including evidence for activation of a TGF-β signaling cascade. The heterodimeric protein clusterin (apolipoprotein J) was also identified as a prominent node in the network. Responsiveness of TGF-β signaling and clusterin gene and protein expression were confirmed independently of the microarray data, verifying some components of the model. This is the first attempt to develop a comprehensive molecular network for histological BPH induced by adrenergic activation. The study also implicated clusterin as a novel biochemical target for therapy.

The patient was a 34-year-old man presenting with the right intra-scrotal painless mass. With a diagnosis of right intrascrotal tumor, the patient underwent right high orchiectomy. The pathological diagnosis of pleomorphic rhabdomyosarcoma arisen from the right spermatic cord was made. Computed tomography revealed a single metastasis in the para-vena cava lymph node. Systemic chemotherapy with vincristine, actinomycin D, plus cyclophosphamide (VAC therapy), and etoposide plus cisplatin (EP therapy) were made according to Intergroup Rhabdomyosarcoma Study (IRS)-IV Regimen 45. But the chemotherapy was ineffective and a retoroperitoneal lymphadenectomy (RPLND) was therefore performed. After 3 months following RPLND, the tumor relapsed in a pelvic lymph node involved in right ureter and ileocaecal valve. Resection of the tumor with ileocaecum was performed and then intraoperative radiotherapy (15 Gy) against the tumor bed was performed to ensure the curative effects. After his recovery, he received a total of 6 courses of systemic chemotherapy consisting of vincristin, ifosphamide, etoposide (IRS-IV Regimen 47). The patient was rigorously followed up for 42 months after the final chemotherapy, with no tumor recurrence.

Objectives: Aquaporins (AQP) function as selective pores allowing water, glycerol and other small solutes to pass through the cell membrane. We previously reported that AQP are expressed in the prostates of both humans and rats. The present study investigated the androgen-dependent expression of AQP9 in the prostate in vivo and in vitro. Methods: Rat ventral prostate tissue specimens and a normal human prostatic epithelial cell line (PNT2) were used. AQP9 messenger ribonucleic acid (mRNA) and protein expression were examined using real-time polymerase chain reaction, Western blotting and immunohistochemical methods. Androgen modulation was achieved by surgical castration, treatment with testosterone propionate (5 µg/g bodyweight) or bicalutamide (20 µg/g bodyweight), and the ribonucleic acid interference (RNAi) method of the androgen receptor (AR). The synthetic small interfering RNA was transfected into PNT2 at a concentration of 10 nM, and the RNAi effect was evaluated using a Western blotting analysis. Results: AQP9 mRNA and protein were expressed in the rat prostate. Surgical castration or bicalutamide treatment significantly decreased their expression. In addition, the treatment with testosterone propionate after castration restored the expression to the level of the controls. An RNAi experiment in PNT2 also decreased the expression. Conclusions: AQP9 expression in the prostate is controlled by androgens.

Objective: Aquaporins (AQPs) function as selective pores allowing water, glycerol and other small solutes to pass through the cell membrane. The present study investigated the expression and localization of AQP3 in the human prostate. Methods: Three human prostate cancer cell lines (DU-145, LNCaP and PC-3) and one normal human prostate cell line (PNT1A) were used in this study. Clinical materials from patients with prostate cancer were also used. AQP3 mRNA and protein expression were examined using reverse transcription-polymerase chain reaction and immunohistochemical methods. Results: Aquaporin 3 mRNA was expressed in normal and cancerous prostate cells. Moreover, the expression of AQP3 mRNA was seen in both normal and cancerous epithelia of human prostate tissues, but not in the mesenchyme. In the normal epithelia of the prostate, localization was limited to cell membranes, particularly the basolateral membranes. However, the expression of AQP3 protein in the cancer epithelia was not observed on the cell membranes. Conclusion: The results suggest that water-channel protein may be involved in the preservation of cellular character in the human prostate and prostate cancer is associated with an alteration of water-transporting mechanisms. Changes in the localization of AQP3 in cancer cells may result from tumorigenesis.

INTRODUCTION AND OBJECTIVES: Fluorodeoxygulcose positron emission tomography/computed tomography (FDG-PET/CT) is becoming useful for diagnosis, staging and prognosis in many cancers. In contrary, the use in urological oncology has been slower to develop because the radiotracer is excreted into the urine. Then, to assess the ability of preoperative FDG-PET/CT to detect upper urinary tract cancers (UUTC), compared with pathological examinations of tissues obtained by ureteroscopic biopsy or split cytologic analysis of urines obtained following retrograde pyelography. METHODS: Clinicopathological records of patients who were examined by FDG-PET/CT were retrospectively reviewed. Sixty-four patients (66 lesions) with clinically suspected UUTC, who were diagnosed by ureteroscopy or nephroureterectomy or partial ureterectomy at our institution from September 2010 to September 2014, were included. The patient cohort consisted of 51 men and 13 women, with a median age of 73 (range 54e92) years. RESULTS: 66 lesions were histologically diagnosed as 59 urothelial carcinomas and 1 clear cell carcinoma and 6 benign lesions. Only 22% of 58 lesions with UUTC had positive voided urine cytology and 45% of 47 lesions had positive split urine cytology. In addition, only 53 % of 15 lesions with UUTC had positive endoscopic biopsy. However, 83% of 60 lesions with UUTC had positive FDG-PET/CT examination. The positive predictive value was 93%. The sensitivities of

金沢大学大学院医学系研究科機能再生学, The clinical significance of hyaluronan (HA) levels in the cerebrospinal fluid (CSF) of patients with spinal tumor (ST) was evaluated in order to clarify whether HA concentrations in the CSF of patients with ST differ according to such factors as the tumor site and histopathological diagnosis. CSF samples were obtained from 40 patients with ST who had undergone myelography and CSF examinations retrospectively. The HA levels were determined using a sandwich-binding protein assay. The total protein (TP) levels were also determined. The HA and TP concentrations in CSF were significantly higher in patients with extramedullary tumor than in patients with intramedullary tumor. There was a significant correlation between HA and TP concentrations in CSF patients with ST. A HA assay for CSF is therefore considered to be potentially useful for estimating the localization of ST. © 2006 World Scientific Publishing Company.

Clinical studies with prostate cancer tissue indicate that alterations in androgen receptor (AR) or c-Met overexpression are associated with androgen-independent progression. We investigated the interaction between AR and c-Met signaling in human prostate cancer cells. Androgen withdrawal or AR-specific small interfering RNA significantly reduced the growth rate while each maneuver induced the expression of c-Met. Knockdown of both AR and c-Met expression markedly inhibited the cell growth. Furthermore, microarray analysis indicated that the activation of c-Met down-regulated the expression of DNA repair-related genes including 8-oxoguanine DNA glycosylase. Exogenous hepatocyte growth factor also induced the production of intracellular reactive oxygen species and resulted in the accumulation of DNA damages. These results suggested that the activation of c-Met signaling may lead to induction of spontaneous mutations or genomic instability, which may lead to the progression of androgen-independent state. Thus, c-Met signaling is utilized for survival and growth under the androgen-depleted condition.

STUDY DESIGN: A case report of an extremely rare malignant spinal tumor successfully treated with total en bloc spondylectomy and chemotherapy. OBJECTIVE: To describe points for consideration when an osteogenic lesion in the spine is diagnosed and treated. SUMMARY OF BACKGROUND DATA: Primary mesenchymal chondrosarcoma in the spine is extremely rare. There were no reports of this tumor being treated with spondylectomy to achieve total surgical resection with a wide margin followed by chemotherapy. METHODS: A 44-year-old female presented with low back pain and left flank pain. Magnetic resonance imaging and computerized tomography showed an osteosclerotic tumor of the lumbar vertebrae. Tc-99m HMDP bone scintigraphy was positive, but thallium-201 scintigraphy and gallium scintigraphy were negative. The patient was diagnosed as having chondrosarcoma based on biopsy findings. RESULTS: To resect the tumor completely, total en bloc spondylectomy for 2 consecutive lumbar vertebrae was performed. However, the postoperative pathologic diagnosis was extremely difficult because the patient was initially suspected to have osteosarcoma, but the final diagnosis was mesenchymal chondrosarcoma. Five years after surgery, there have not been any signs of local recurrence or distant metastasis, and the patient has remained continuously disease free. CONCLUSIONS: To our knowledge, we reported the first case of mesenchymal chondrosarcoma occurring from the lumbar spine treated with total en bloc spondylectomy and chemotherapy. Successful radical resection of the tumor could be accomplished. Although the effect of chemotherapy on the final results could not be clearly determined, considering that at least continuous disease-free survival was achieved, it is highly likely that chemotherapy contributed to the favorable results.

It is well known that the coagulation factor XIII (FXIII) is deeply involved in wound healing and tissue regeneration. We measured the plasma levels of FXIII (FXIII levels) before and after surgery in 80 patients with bone tumors [benign tumors in 30 patients (BT group), malignant tumors in 30 patients (MT group), and carcinoma metastasis in 20 patients (CM group)] and in the control group which consisted of 30 patients with osteoarthritis of the hip joint who had underwent total hip arthroplasty (THA). The numbers of patients whose preoperative FXIII level was below the lower limit of normal were as follows: 0 (0%) among 30 patients in the control group; 7 (23%) among 30 patients in the BT group; 12 (40%) among 30 patients in the MT group; and 9 (45%) among 20 patients in the CM group. Among patients with bone tumors examined in this study, patients with osteosarcoma who underwent chemotherapy showed a significant decrease in FXIII levels (p < 0.01). We indicated not only FXIII levels in patients with bone tumors, but also the natural time-course of FXIII levels in patients who underwent orthpaedic surgery of THA at the first time.

An 80-year-old man presented to the internist with fever, fatigue and leukocytosis up to 66.8 x 10(3)/microl. Although a chronic myelogenous leukemia was initially suspected, he was diagnosed as metastatic bone marrow tumor with bone marrow necrosis from primary prostate cancer on the basis of the clinical and pathological findings. The serum concentrations of IL-6 and TNF-alpha were mildly elevated to 65.0 pg/ml and, 54.0 pg/ml respectively. It is probable that these humoral factors were partially responsible for the leukemoid reaction although other factors induced by the bone marrow necrosis with bone marrow metastasis of prostate cancer are also likely involved.

Urachal cancers are rare bladder cancers and almost always adenocarcinomas. Surgical resection is the only standard treatment, and no effective chemotherapeutic regimen has been established. We herein report a case of urachal cancer that recurred 2 years after tumor resection and partial cystectomy. The patient was successfully treated with surgery and modified FOLFOX6 (mFOLFOX6) chemotherapy. A 60-year-old female patient diagnosed with urachal cancer underwent tumor resection and partial cystectomy at another hospital. The initial clinical and pathological stage was unclear. Two years later, an intrapelvic mass that had invaded the uterus and right ovary and multiple lung metastases were detected on follow-up computed tomography. The intrapelvic tumor, entire uterus, vault of the vagina, and right ovary were completely excised at another different hospital. The patient was referred to our department for further management of multiple lung metastases. A combined chemotherapeutic regimen with mFOLFOX6 (85 mg/m2 oxaliplatin and 200 mg/m2 leucovorin administered as 2-h infusions on day 1, followed by a 400 mg/m2 5-fluorouracil bolus and 46-h infusion of 2400 mg/m2 over days 1 and 2) was administered every 2 weeks as one cycle. After six cycles of treatment, CT revealed that the lung metastases had completely disappeared. In total, the patient underwent 19 cycles of the regimen. At the time of this report, the patient has been free from recurrence for 34 months. The successful treatment observed in the present case sheds light on a possible curative treatment for this disease using a multimodal treatment strategy, particularly surgical excision of intrapelvic tumors and the administration of mFOLFOX for distant metastases.

Gastroduodenal ulcers in renal transplant recipients are usually originated from excessive acid secretion or infection of Helicobacter pyroli. Herein, we report a case of cytomegalovirus (CMV)--induced gastric ulcer following cadaveric renal transplantation. The patient was a 48-year-old man with chronic renal failure and received cadaveric renal transplantation. A month later, he had epigastralgia without CMV-positive antigenemia and received gastrointestinal fiberscopy. Endoscopically, gastric ulcer was identified. Histological findings revealed conspicious nuclear enlargement of the non-epithelial cells in the ulcer bed, which indicated CMV infection. The patient was treated with ganciclovir for 2 weeks and the symptom was relieved. He discharged with a good renal function on day 75 posttransplant. CMV infection plays an important role in gastric ulcer after renal transplantation. Antigenemia assay dose not seem feasible for the detection of CMV-induced gastric ulcer.

Diabetic nephropathy ensues from events involving earliest changes in the glomeruli and podocytes, followed by accumulation of extracellular matrix in the mesangium. Postulated mechanisms include roles for vascular endothelial growth factor (VEGF), produced by podocytes and contributing to enhanced excretion of urinary albumin and recruitment/activation of inflammatory cells, and transforming growth factor-beta (TGF-beta), elicited largely from mesangial cells and driving production of extracellular matrix. RAGE, a receptor for advanced glycation endproducts (AGEs) and S100/calgranulins, displays enhanced expression in podocytes of genetically diabetic db/db mice by age 13 weeks. RAGE-bearing podocytes express high levels of VEGF by this time, in parallel with enhanced recruitment of mononuclear phagocytes to the glomeruli; events prevented by blockade of RAGE. By age 27 weeks, soluble RAGE-treated db/db mice displayed diminished albuminuria and glomerulosclerosis, and improved renal function. Diabetic homozygous RAGE null mice failed to develop significantly increased mesangial matrix expansion or thickening of the glomerular basement membrane. We propose that activation of RAGE contributes to expression of VEGF and enhanced attraction/activation of inflammatory cells in the diabetic glomerulus, thereby setting the stage for mesangial activation and TGF-beta production; processes which converge to cause albuminuria and glomerulosclerosis.

The effect of castration on apoptosis in the mouse epididymis during postnatal development was examined. The weight of the epididymis slowly increased from day 0 (day of birth) to day 20 after birth, followed by a rapid increase thereafter. Castration on days 0, 5, 10, 20, 30, 40 and 60 increased apoptotic indices (percentages of apoptotic cells) of epithelia of the caput (head), corpus (body), and cauda (tail) epididymis, their apoptotic indices reaching maximal levels on day 2 after castration with the exception of a maximal apoptotic index on day 4 in the tail after castration on day 60. The maximal levels of apoptotic indices of the head, body and tail after castration on days 0, 5, 10 and 20 were significantly lower than those after castration on days 40 and 60. DNAs extracted from the epididymides 2 days after castration on days 0, 5, 10 and 60 showed a ladder pattern on agarose gel electrophoresis, which is a characteristic of apoptosis. When testosterone propionate (10 microg/g body weight) was injected twice a day into mice which had been castrated on day 10, 30 or 60, the increases in apoptotic indices of the head, body and tail of the epididymis were completely inhibited. The weights of the paired epididymides 6 days after castration on days 0, 5, 10, 20, 30, 40 and 60 were significantly lower than those of sham-operated mice, indicating the secretion of androgen by the testes from birth to adulthood. The present results indicated that androgen deprivation caused by castration induces apoptosis in the epithelium of the epididymis of mice from birth to adulthood, and suggested that a proportion of epithelial cells, the survival of which is dependent on the testes, is smaller in the epididymides during a slow growth stage than in the epididymides after this stage.

The purpose of this study was to assess the ability of fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) to detect upper urinary tract urothelial carcinomas (UTUC) compared with pathological examination of tissues obtained by ureteroscopic biopsy and split cytologic analysis of urine obtained after retrograde pyelography. Clinicopathological records of patients at our institution were retrospectively reviewed. Fifty patients with clinically suspected UTUC, who were histologically diagnosed by nephroureterectomy, partial ureterectomy, or endoscopic biopsy, were included. The patient cohort included 42 men and 8 women, with a median age of 73 (range 54–92) years. Only 27 % of 49 patients with UTUC had positive voided urine cytology, and 33 % of 40 patients had positive split urine cytology. In addition, 40 % of 10 patients had a positive endoscopic biopsy. However, 83 % of 48 patients with UTUC had positive results from FDG-PET/CT examination. The positive predictive value of FDG-PET/CT was 95 %. There were no correlations between sensitivity and tumor stage or tumor grade. Sensitivity of FDG-PET/CT for patients with and without diabetes mellitus was 60 and 89 %, respectively. These preliminary results from a small number of patients revealed that FDG-PET/CT enabled effective detection of UTUC.

PURPOSE Adrenocortical carcinoma (ACC) is a rare condition associated with poor prognosis. This study aimed to evaluate the clinicopathologic characteristics and prognosis of 7 patients with ACC. PATIENTS AND METHODS The clinicopathologic characteristics, treatment, and survival of 7 patients with pathologically confirmed ACC treated at our institution between January 2002 and December 2012 were retrospectively examined. RESULTS The study cohort comprised 4 male and 3 female patients (median age at diagnosis, 63 years [range, 36-71 years]). The median tumor size was 7.0 cm (range, 4.0-13.0 cm), and the median follow-up duration was 22 months (range, 9-107 months). One patient had stage I ACC, 4 had stage III, and 2 showed metastasis. The patient with stage I disease underwent laparoscopic adrenorectomy and those with stage III disease underwent adrenorectomy with the excision of adjacent organs. Four of these 5 patients are alive without recurrence at a median of 55 months (range, 22-107 months) after surgery. Of the 2 patients with metastases, 1 received combined chemotherapy with etoposide, adriamycin, and cisplatin plus mitotane without surgical resection but died 19 months later, and the other, with a solitary lung metastasis, underwent adrenorectomy and metastatectomy followed by adjuvant treatment with mitotane and is alive without recurrence at 9 months after treatment. The 3-year cause-specific survival rate was 56%. CONCLUSIONS Patients with advanced-stage tumors showed long-term survival with complete tumor resection at diagnosis; hence, this seems to be most beneficial treatment option for patients with ACC.

Abstract —Advanced glycation end products (AGEs) and their cell surface receptor, RAGE, have been implicated in the pathogenesis of diabetic complications. Here, we studied the role of RAGE and expression of its proinflammatory ligands, EN-RAGEs (S100/calgranulins), in inflammatory events mediating cellular activation in diabetic tissue. Apolipoprotein E–null mice were rendered diabetic with streptozotocin at 6 weeks of age. Compared with nondiabetic aortas and kidneys, diabetic aortas and kidneys displayed increased expression of RAGE, EN-RAGEs, and 2 key markers of vascular inflammation, vascular cell adhesion molecule (VCAM)-1 and tissue factor. Administration of soluble RAGE, the extracellular domain of the receptor, or vehicle to diabetic mice for 6 weeks suppressed levels of VCAM-1 and tissue factor in the aorta, in parallel with decreased expression of RAGE and EN-RAGEs. Diabetic kidney demonstrated increased numbers of EN-RAGE–expressing inflammatory cells infiltrating the glomerulus and enhanced mRNA for transforming growth factor-β, fibronectin, and α 1 (IV) collagen. In mice treated with soluble RAGE, the numbers of infiltrating inflammatory cells and mRNA levels for these glomerular cytokines and components of extracellular matrix were decreased. These data suggest that activation of RAGE primes cells targeted for perturbation in diabetic tissues by the induction of proinflammatory mediators.

Mesenchymal-epithelial interactions are extremely important during growth and development and in the functional cytodifferentiation of male sex accessory organs. Interactions between mesenchyme and epithelium occur mainly through a paracrine action that is mediated by various growth factors. The role of androgens is very important for these organs and the androgenic effect is mediated by paracrine interactions. A number of growth factors have been studied in prostate and seminal vesicles from mice, rats, and humans because they are potent mediators of cellular proliferation, differentiation, and death. This review provides an overview of current knowledge about growth factors involved in the development of male sex accessory organs, with particular emphasis on the prostate.

Advanced glycation end products (AGE) contribute to diabetic tissue injury by two major mechanisms, i.e., the alteration of extracellular matrix architecture through nonenzymatic glycation, with formation of protein crosslinks, and the modulation of cellular functions through interactions with specific cell surface receptors, the best characterized of which is the receptor for AGE (RAGE). Recent evidence suggests that the AGE-RAGE interaction may also be promoted by inflammatory processes and oxidative cellular injury. To characterize the distributions of AGE and RAGE in diabetic kidneys and to determine their specificity for diabetic nephropathy, an immunohistochemical analysis of renal biopsies from patients with diabetic nephropathy (n = 26), hypertensive nephrosclerosis (n = 7), idiopathic focal segmental glomerulosclerosis (n = 11), focal sclerosis secondary to obesity (n = 7), and lupus nephritis (n = 11) and from normal control subjects (n = 2) was performed, using affinity-purified antibodies raised to RAGE and two subclasses of AGE, i.e., N(epsilon)-(carboxymethyl)-lysine (CML) and pentosidine (PENT). AGE were detected equally in diffuse and nodular diabetic nephropathy. CML was the major AGE detected in diabetic mesangium (96%), glomerular basement membranes (GBM) (42%), tubular basement membranes (85%), and vessel walls (96%). In diabetic nephropathy, PENT was preferentially located in interstitial collagen (90%) and was less consistently observed in vessel walls (54%), mesangium (77%), GBM (4%), and tubular basement membranes (31%). RAGE was expressed on normal podocytes and was upregulated in diabetic nephropathy. The restriction of RAGE mRNA expression to glomeruli was confirmed by reverse transcription-PCR analysis of microdissected renal tissue compartments. The extent of mesangial and GBM immunoreactivity for CML, but not PENT, was correlated with the severity of diabetic glomerulosclerosis, as assessed pathologically. CML and PENT were also identified in areas of glomerulosclerosis and arteriosclerosis in idiopathic and secondary focal segmental glomerulosclerosis, hypertensive nephrosclerosis, and lupus nephritis. In active lupus nephritis, CML and PENT were detected in the proliferative glomerular tufts and crescents. In conclusion, CML is a major AGE in renal basement membranes in diabetic nephropathy, and its accumulation involves upregulation of RAGE on podocytes. AGE are also accumulated in acute inflammatory glomerulonephritis secondary to systemic lupus erythematosus, possibly via enzymatic oxidation of glomerular matrix proteins.

We report the occurrence of congenital nephrotic-range proteinuria secondary to focal segmental glomerulosclerosis in an infant with epidermolysis bullosa and pyloric atresia. A homozygous missense mutation, R1281W, in exon 31 of the beta4 integrin gene, ITGB4, was identified. By immunofluorescence, beta4 integrin expression was reduced in both dermal keratinocytes and glomerular podocytes. This is the first demonstration of beta4 integrin expression in human glomeruli. We postulate a role for altered beta4 integrin function in the mediation of the glomerular permeability defect.

The receptor for advanced glycation end products (RAGE), a multi-ligand member of the immunoglobulin superfamily of cell surface molecules, interacts with distinct molecules implicated in homeostasis, development and inflammation, and certain diseases such as diabetes and Alzheimer's disease. Engagement of RAGE by a ligand triggers activation of key cell signalling pathways, such as p21ras, MAP kinases, NF-kappaB and cdc42/rac, thereby reprogramming cellular properties. RAGE is a central cell surface receptor for amphoterin, a polypeptide linked to outgrowth of cultured cortical neurons derived from developing brain. Indeed, the co-localization of RAGE and amphoterin at the leading edge of advancing neurites indicated their potential contribution to cellular migration, and in pathologies such as tumour invasion. Here we demonstrate that blockade of RAGE-amphoterin decreased growth and metastases of both implanted tumours and tumours developing spontaneously in susceptible mice. Inhibition of the RAGE-amphoterin interaction suppressed activation of p44/p42, p38 and SAP/JNK MAP kinases; molecular effector mechanisms importantly linked to tumour proliferation, invasion and expression of matrix metalloproteinases.

The rat ventral prostate gland is a model tissue to study the effects of androgenic steroids on prostate cells. Recent reports suggest that the prostatic vascular system is a primary target of androgen action in this tissue. In order to better understand how the vascular system of the ventral prostate supports the tissue in an androgenically normal adult male rat we utilized a variety of microscopic imaging techniques to more fully characterize its structural anatomy and its interaction with other prostatic cell types. Vascular corrosion casts were produced from the mature ventral prostate glands of rats. These casts were analyzed by scanning electron microscopy (SEM) to describe gross and fine details of the prostate vascular anatomy. Fixed thin sections of ventral prostates were immunostained with antiFactor XIII and analyzed by light microscopy for the presence of capillary elements within the prostatic glands. Other sections were directly analyzed by transmission electron microscopy (TEM) to describe the anatomical relationship between the capillaries and the prostatic ducts and their associated glands. The rat ventral prostate is supplied with blood by branches of the inferior vesical artery which enters the apex of the tissue from the base of the urinary bladder. Visualization of the prostatic vascular network under SEM shows that this major vessel is found on the posterior-medial surface of the tissue (closest to the bladder). This surface also has numerous serpentine vessels that appear to facilitate a stable blood supply to the prostate in accommodation of urinary bladder distension. Examination of the opposing surface of the casts allowed a crude description of the structure of the prostatic ductal system with the distal tips of the ducts (containing the prostate glands) oriented towards the anterior-lateral surface of the tissue. On this surface, one can discern a series of adjacent basket-shaped vascular structures of distributing arterioles that supply a dense complex of fine capillaries to the glands. Analysis of the interface of the prostatic ductal system with its vascular elements by light microscopy and TEM shows that some capillaries lie immediately adjacent to the basement membranes of the glands while others can be found interspersed within the myofibroblast layer surrounding the ducts and glands. Veins accompany the arteries and combine with the superior vesical before entering the common iliac vein. This study gives a comprehensive and detailed view of the microvasculature of the rat ventral prostate gland. The findings here will provide the basis for future experiments to evaluate how the ventral prostate vascular system changes in response to androgenic manipulation and to other pathological conditions. Anat Rec 256:403–411, 1999. © 1999 Wiley-Liss, Inc.

Background: Little is known about the mechanisms underlying recovery from hydronephrosis. Using a rat kidney model, we investigated the possible contribution of growth factors during regeneration after hydronephrosis. Methods: After the unilateral ureteral obstruction for 7 days, the obstruction was released by ureterocystostomy. Epidermal growth factor (EGF), hepatocyte growth factor (HGF) and transforming growth factor-β1 (TGF-β1) were studied using immunohistochemistry and semiquantitative reverse transcriptase–polymerase chain reaction (RT-PCR). Results: The immunoreaction of EGF in the medulla of both obstructed and contralateral kidneys increased after releasing the obstruction. The release of ureteral obstruction brought further increased immunoreaction of HGF to both the cortex and the medulla of the contralateral kidney. The immunoreaction of TGF-β1 also increased in the interstitium especially around the blood vessels in the post-obstructed kidney. The expression of HGF and EGF mRNA in both kidneys and TGF-β1 mRNA in the obstructed kidney were increased after releasing the obstruction. Conclusions: These results suggest that various growth factors may be involved in the post-obstructive tubular recovery and interstitial damage in the rat kidney.

Background: Information concerning varicocele is limited in spite of its clinical importance. This study intends to clarify anatomical features of varicocele. Methods: Spermatic veins were obtained from varicocele patients at surgery. Areas of two distinct portions (extracellular matrix and smooth muscle) were measured by a micro computer imaging device image analysis system on histologic slides with Masson trichrome stain. In addition, we evaluated the shape and arrangement of the smooth muscle fibers by a scanning electron microscope (SEM). Results: Masson trichrome stain revealed that grade of varicocele partly correlated with thickness of the wall and with amount of the smooth muscle fibers. In the patients with grade 3 varicocele, SEM examination showed marked hypertrophy of longitudinal smooth muscle fibers, which were discriminated clearly from the inner circular fibers. Conclusions: This study indicated that morphological changes occurred in the muscle layers of varicoceles in the worst grade of varicocele. These structural changes which result from abnormal venous blood circulation may cause development of varicocele, resulting in male infertility.

In the testis, several types of heat shock proteins (HSPs) have been identified and characterized, although the cellular basis of the HSPs remains elusive. In the present study, alterations in the cellular localization of HSPs, including HSP 25, 60, 70, and 90, were studied during the developing and degenerating periods in the rat testis using immunohistochemistry and Western blotting. HSP25 was expressed in neither germ cells nor somatic cells on all days examined. In contrast, HSP 60 was expressed in Leydig cells during neonatal and prepuberty periods, and only in spermatogonia and primary spermatocytes after puberty. HSPs 70 and 90 were expressed in germ cells, Sertoli cells, and Leydig cells during neonatal and early developing testes, and in spermatocytes and round spermatids after puberty. Besides, there was faint expression of HSP 90 protein in spermatogonia in this period. In the degenerative condition, all HSP proteins were markedly expressed in germ cells after surgery. It would appear that HSPs play roles in unique homeostasis in testes.

Background: information concerning the mechanisms underlying recovery from hydronephrosis is limited. The frequency of apoptosis during healing from hydronephrosis was studied using a rat kidney model. Methods: The presence of apoptosis was studied using an in situ DNA 3 end labeling method, electron microscopy, and agarose gel electrophoresis. Results: The degree of apoptosis in both the medulla and cortex gradually increased during ureteral obstruction as shown by in situ DNA 3 end labeling. Release of the ureteral obstruction resulted in a further increase in the degree of apoptosis in the medulla and cortex. The increase in apoptosis in the medulla was transient and lasted for only 4 days following release, while that in the cortex continued for at least 3 weeks. Apoptosis in the glomerulus was not observed. Electron microscopy revealed cells with aggregated chromatin in compact granular masses that abutted the nuclear membrane. Following release of ureteral obstruction, DNA fragmentation characteristic of apoptosis was visible on agarose gel electrophoresis. Conclusion: These results suggest that apoptosis is involved in post-obstructive tubular damage in the rat kidney.

Objective: To evaluate biochemical recurrence-free survival (b-RFS) in patients with Gleason score 7 prostate cancers treated with external beam radiotherapy at Ehime University Hospital. Materials and Methods: Between January 2003 and October 2009, 63 patients with Gleason score 7 prostate cancers were treated with three-dimensional conformal radiotherapy (3D-CRT) at our institute. Of the 63 patients analyzed, 41 and 22 had a primary Gleason pattern of 3 and 4 carcinoma, respectively. Neoadjuvant hormonal therapy had been given to 37 patients (59%) for 6 months prior to radiotherapy. The American Society for Therapeutic Radiology and Oncology Phoenix consensus definition was used to determine the b-RFS after treatment. Results: The overall b-RFS rate at 5 year was 71% and 77% for Gleason score 3+4 and 4+3 prostate cancer, respectively. The overall b-RFS at 5 year was 59% and 86% in Gleason score 3+4 patients with and without neoadjuvant hormonal therapy for 6 months, respectively. Conclusions: Our results indicate that the 5 year b-RFS outcome with 3D-CRT is not dependent on Gleason score 3+4 versus 4+3 histological features, or on neoadjuvant hormonal therapy for 6 months in patients with a Gleason score of 3+4.

Here, we report a case of primary intracranial tumor in a chronic hemodialysis patient in which neurosurgery was successful. A 50-year-old man who had been on hemodialysis for 4 years was admitted to our hospital with general fatigue. Neurological examination on admission revealed mild restless. Computed tomography and magnetic resonance imaging performed on admission revealed a large (55 mm×40 mm) tumor mass in contact with the falx. The size of this tumor rapidly increased over the next month. 201Thallium-chloride single photon emission computed tomography revealed abnormal uptake in the same location as the lesion. This suggested a malignant brain tumor and surgical excision was scheduled. Two weeks prior to surgery, frequent hemodialysis was performed using nafamostat mesilate instead of heparin to prevent bleeding and to maintain electrolyte balance, and red cell concentrates and erythropoietin were administered for the improvement of anemia. A triple lumen catheter was inserted in the right internal jugular vein in preparation for emergency continuous hemodiafiltration to maintain homeostasis of circulatory dynamics. Surgery was completed without incident and the tumor was resected totally. During surgery, cerebral edema was well controlled by hyperventilation and a slightly upturned head position. Histopathological examination of the specimen confirmed atypical meningioma. Continuous hemodiafiltration was performed for 24 hours after surgery, and hemodialysis was initiated on the third day after surgery. The postoperative course was uneventful. Three weeks after surgery, the patient was discharged with no neurological deficit and resumed his daily life on maintenance hemodialysis.

Background: To improve the prognosis of patients with urachal cancer and establish an effective chemotherapeutic regimen for distant metastases. Methods: We conducted a retrospective study to evaluate the efficacy and safety of a modified combination of 5-fluorouracil, leucovorin and oxaliplatin (mFOLFOX6) therapy in patients with metastatic urachal cancer. Results: Five patients were treated with mFOLFOX6. Their median age was 65 years (range 41-80). The median follow-up time was 42 months (range 18-46). Two of the 5 patients (40%) showed an objective response: 1 achieved a clinically complete response and 1 a partial response. The grade 3/4 toxicity associated with this regimen was primarily neutropenia, but febrile neutropenia was not observed. Oxaliplatin treatment was discontinued because of a grade 2 allergic reaction in 1 patient. Grade 2 peripheral sensory neuropathy caused by oxaliplatin was observed in 2 patients, and the OPTIMOX (stop and go) approach had to be adopted. Conclusions: mFOLFOX6 appears to be effective for the treatment of metastatic urachal cancer.

An 8-year-old boy was admitted to Ehime University Hospital, Ehime, Japan, for the further investigation of a 5-month episode of gross hematuria accompanied by lower abdominal pain. Magnetic resonance imaging revealed a solid tumor measuring 3 cm in diameter of the bladder wall. Cystoscopy demonstrated a red, wide-based, nodular tumor situated on the dome of the bladder. Histological examination of tissue taken at hot biopsy showed fibrolipoma. In consideration of potential malignancy, a partial cystectomy was carried out after informed consent was given. Histological examination of the resected specimen showed it to be cavernous hemangioma.

The effects of suramin on the growth of seminal vesicles (SVs) of neonatal mice were investigated in vitro. SVs from 0-day-old male mice were cultured in serum-free chemically defined medium supplemented with 5 alpha-dihydrotestosterone (DHT, 10(-8) M) and insulin (10 micrograms/ml), alone and in combination. Prior to culture, SVs from 0-day-old mice had no epithelial branches. SVs cultured in medium with DHT formed numerous epithelial branches, while epithelial branching did not occur in SVs cultured without DHT. The addition of suramin (0.2 mM) to medium containing DHT inhibited the formation of epithelial branches almost completely. Removal of suramin from the medium on days 2, 4, and 6 of culture initiated the formation of epithelial branches. Suramin (0.2 mM) reversibly decreased 3H-thymidine-labeling indices (3H-LI) of both epithelium and mesenchyme of SVs cultured in medium with DHT plus insulin or DHT alone during 8 days of culture. Suramin also decreased 3H-LI of both epithelium and mesenchyme of SVs cultured in medium with insulin alone. The present study indicates that suramin reversibly inhibits not only androgen-dependent but also androgen-independent growth and ductal branching morphogenesis of neonatal mouse SVs.

This was a retrospective study to evaluate the activity and toxicity of a combined chemotherapeutic regimen of gemcitabine and carboplatin (GCa) in patients with metastatic urothelial carcinomas (UCs) with special regard to patients with highly impaired renal function.Eleven patients whose creatinine clearance was 30 ml/min or under and who had been diagnosed with metastatic UC were treated with GCa. The patient cohort comprised 4 males and 7 females, with a median age of 74 (range 67-84) years. The median follow-up was 19 (range 1-58) months.Five of the 11 patients (45%) showed an objective response, with 2 achieving a clinically complete response and 3 a partial response with GCa. The grade 3/4 toxicity of the regimen was primarily hematological, including anemia (55%), neutropenia (45%), and thrombocytopenia (45%). Four patients (36%) could not complete the treatment in total. Grade 3 pneumonitis was found in one patient, and the treatment was terminated. Grade 4 febrile neutropenia occurred in the patient on hemodialysis, and the patient was forced to discontinue the chemotherapy. Another 2 patients also called off the treatment due to a pulmonary adverse event and an elevation of serum creatinine, respectively.GCa appears to be effective for the treatment of metastatic UCs in patients with impaired renal function, but it is necessary to pay attention to the occurrence of severe adverse events.

Cisplatin is currently the most effective antitumor agent available against bladder cancer. However, a majority of patients eventually relapse with cisplatin-resistant disease. Chemoresistance thus remains a major obstacle in bladder cancer therapy. To clarify the molecular mechanisms underlying cisplatin resistance in bladder cancer, we established a cisplatin-resistant subline from the human bladder cancer cell line HT1376 (HT1376-CisR), and conducted large-scale analyses of the expressed proteins using two-dimensional (2D) gel electrophoresis coupled with mass spectrometry (MS). Comparative proteomic analysis of HT1376 and HT1376-CisR cells revealed 36 differentially expressed proteins, wherein 21 proteins were upregulated and 15 were downregulated in HT1376-CisR cells. Among the differentially regulated proteins, adseverin (SCIN), a calcium-dependent actin-binding protein, was overexpressed (4-fold upregulation) in HT1376-CisR, with the increase being more prominent in the mitochondrial fraction than in the cytosol fraction. SCIN mRNA knockdown significantly reduced cell proliferation with mitochondria-mediated apoptosis in HT1376-CisR cells. Immunoprecipitation analysis revealed voltage-dependent anion channels (VDACs) to be bound to SCIN in the mitochondrial fraction. Our results suggest that the VDAC-SCIN interaction may inhibit mitochondria-mediated apoptosis in cisplatin-resistant cells. Targeting the VDAC-SCIN interaction may offer a new therapeutic strategy for cisplatin-resistant bladder cancer.