Suppression of ERß signaling via ERß knockout or antagonist protects against bladder cancer development.

Epidemiological studies showed that women have a lower bladder cancer (BCa) incidence, yet higher muscle-invasive rates than men, suggesting that estrogen and the estrogen receptors, estrogen receptor alpha (ERα) and estrogen receptor beta (ERß), may play critical roles in BCa progression. Using in vitro cell lines and an in vivo carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced mouse BCa model, we found that ERß plays a positive role in promoting BCa progression. Knockdown of ERß with ERß-shRNA in ERß-positive human BCa J82, 647v and T24 cell lines led to suppressed cell growth and invasion. Mice lacking ERß have less cancer incidence with reduced expression of the proliferation marker Ki67 in BBN-induced BCa. Consistently, our results show that non-malignant urothelial cells with ERß knockdown are more resistant to carcinogen-induced malignant transformation. Mechanism dissection found that targeting ERß suppressed the expression of minichromosome maintenance complex component 5 (MCM5), a DNA replication licensing factor that is involved in tumor cell growth. Restoring MCM5 expression can partially reverse ERß knockdown-mediated growth reduction. Supportively, treating cells with the ERß-specific antagonist, 4-[2-Phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP), reduced BCa cell growth and invasion, as well as MCM5 expression. Furthermore, we provide the first evidence that BCa burden and mortality can be controlled by PHTPP treatment in the carcinogen-induced BCa model. Together, these results demonstrate that ERß could play positive roles in promoting BCa progression via MCM5 regulation. Targeting ERß through ERß-shRNA, PHTPP or via downstream targets, such as MCM5, could serve as potential therapeutic approaches to battle BCa.

ASC-J9 suppresses castration-resistant prostate cancer growth through degradation of full-length and splice variant androgen receptors.

Early studies suggested androgen receptor (AR) splice variants might contribute to the progression of prostate cancer (PCa) into castration resistance. However, the therapeutic strategy to target these AR splice variants still remains unresolved. Through tissue survey of tumors from the same patients before and after castration resistance, we found that the expression of AR3, a major AR splice variant that lacks the AR ligand-binding domain, was substantially increased after castration resistance development. The currently used antiandrogen, Casodex, showed little growth suppression in CWR22Rv1 cells. Importantly, we found that AR degradation enhancer ASC-J9 could degrade both full-length (fAR) and AR3 in CWR22Rv1 cells as well as in C4-2 and C81 cells with addition of AR3. The consequences of such degradation of both fAR and AR3 might then result in the inhibition of AR transcriptional activity and cell growth in vitro. More importantly, suppression of AR3 specifically by short-hairpin AR3 or degradation of AR3 by ASC-J9 resulted in suppression of AR transcriptional activity and cell growth in CWR22Rv1-fARKD (fAR knockdown) cells in which DHT failed to induce, suggesting the importance of targeting AR3. Finally, we demonstrated the in vivo therapeutic effects of ASC-J9 by showing the inhibition of PCa growth using the xenografted model of CWR22Rv1 cells orthotopically implanted into castrated nude mice with undetectable serum testosterone. These results suggested that targeting both fAR- and AR3-mediated PCa growth by ASC-J9 may represent the novel therapeutic approach to suppress castration-resistant PCa. Successful clinical trials targeting both fAR and AR3 may help us to battle castration-resistant PCa in the future.

Superhigh-magnetization nanocarrier as a doxorubicin delivery platform for magnetic targeting therapy.

The aim of this study describes the creation of superhigh-magnetization nanocarriers (SHMNCs) comprised of a magnetic Fe(3)O(4) (SHMNPs) core and a shell of aqueous stable self-doped poly[N-(1-one-butyric acid)]aniline (SPAnH), which have a high drug loading capacity (∼27.1 wt%) of doxorubicin (DOX). The SHMNCs display superparamagnetic property with a magnetization of 89.7 emu/g greater than that of Resovist (a commercial contrast agent used for magnetic resonance imaging; 73.7 emu/g). Conjugating the anticancer drug DOX to these nanocarriers enhances the drug's thermal stability and maximizes the efficiency with which it is delivered by magnetic targeting (MT) therapy to MGH-U1 bladder cancer cells, in part by avoiding the effects of p-glycoprotein (P-gp) pumps to enhance the intracellular concentration of DOX. The high R2 relaxivity (434.7 mM(-1)s(-1)) of SHMNCs not only be a most effective MT carrier of chemotherapeutic agent but be an excellent contrast agent of MRI, allowing the assessment of the distribution and concentration of DOX in various tissues and organs. This advanced drug delivery system promises to provide more effective MT therapy and tumor treatment using lower therapeutic doses and potentially reducing the side effects of cardiotoxicity caused by DOX.

Prognostic value of TNM stage and tumor necrosis for renal cell carcinoma.

Our objective was to assess the value of tumor necrosis and other factors for predicting the outcome of renal cell carcinoma (RCC). Our study comprised 328 RCC patients who were surgically treated at this hospital between 2001 and 2006. The five-year survival data was analyzed using a Kaplan-Meier statistical analysis. The prognostic factors were evaluated with a univariate analysis using a log-rank test and multivariate analysis using the Cox proportional hazards regression method. The mean follow-up period for these patients was 46.5 months (median 45.2 months). The univariate analysis revealed that age, tumor stage, TNM stage, grade, tumor necrosis, and histological type were statistically significant prognostic factors. The multivariate analysis showed that the TNM stage and tumor necrosis were the most important predictive factors in the patients' overall survival. In the TNM stage with and without tumor necrosis, the five-year overall survival rates in stages I+II were 80.5% and 89.2%, respectively (p=0.115), where as the five-year survival rates in stages III+IV were 32.7% and 84.0%, respectively (p<0.001). Collectively, our present data revealed that tumor necrosis was an important predictive factor for survival in advanced stage RCC. In conclusion, both the TNM stage and tumor necrosis provided the most important prognostic factors of survival in RCC. Tumor necrosis proved to be a poor prognostic factor in advanced RCCs.

Epstein-Barr virus DNA load in tumour tissues correlates with poor differentiation status in non-muscle invasive urothelial carcinomas.

OBJECTIVE: To detect the correlation between the clinical staging, grading and genomic Epstein-Barr virus (EBV) viral numbers in tumour tissues of urothelial carcinoma patients. PATIENTS AND METHODS: From June 2004 to May 2008, 60 urothelial carcinoma patients (50 cases of bladder carcinoma and 10 of upper tract urothelial carcinoma (UTUC) were enrolled in the study. Eight patients who underwent transurethral resection of prostate for prostate hyperplasia and two patients receiving nephrectomy for non-function kidney were used as normal controls. The EBV viral copy numbers in genomic DNA were evaluated using a real-time PCR-based study. The BamHI W region of the Namalwa cell line was constructed to the plasmid clone and was used as standard curve for absolute quantitative PCR (Q-PCR). RESULTS: Epstein-Barr virus DNA was detected in 56% (28/50) and 60% (6/10) of the bladder and UTUC patients, respectively. The EBV DNA could not be detected in the normal control group. By pooling the UTUC and bladder patients in stage Ta,T1, the high copy number in fixed genomic DNA amount (100 ng/20 µL) was correlated with the high grading in stage Ta,T1 urothelial carcinoma (P = 0.014). The overall grading was not statistically associated with EBV copy number (P = 0.25). Although the copy numbers between paired tumour and normal tissues were not statistically different (P= 0.169), there were more copies of EBV in the normal tissues adjacent to the tumours than in those free from urothelial carcinoma. There was no significant difference between recurrence of non-muscle invasive bladder cancer and the presence of EBV (P > 0.05). CONCLUSIONS: Epstein-Barr virus DNA could be detected in the genome of the urothelial carcinoma specimens. The poor differentiation status was correlated with the high load of the EBV genome in non-muscle invasive urothelial carcinoma. However, recurrence-free survival was not greater in EBV-positive patients than in EBV-negative patients.

Epstein-Barr virus-infected cell line TCC36B derived from B lymphocytes infiltrating renal pelvis urothelial carcinoma.

UNLABELLED: This study reports an initial analysis of an EBV-infected B cell line (TCC36B), established from an urothelial carcinoma (UC) lesion of the renal pelvis. MATERIALS AND METHODS: Cytofluorometric and G-banding analyses were performed for phenotyping and cytogenetics. PCR was used to detect EBV DNA, and sequence analysis to investigate mutations and deletions of the latent membrane protein (LMP)-1 gene of EBV. RESULTS: TCC36B cells proliferated in vitro and showed positivity for surface CD19, CD20, HLA-DR and IgG(λ), indicating that they belong to B-cells. Cytogenetic analysis showed 46,XX with a unique clonal abnormality of dup(2)(p13p25). EBV DNA was detected in TCC36B cells. Sequence analysis revealed a 30-bp deletion and 7 point mutations on the LMP-1 gene in TCC36B cells. CONCLUSION: These results suggest the involvement of an EBV variant in the pathogenesis of UC. This cell line should thus facilitate further investigations on the aetiological role of EBV in urothelial cancer.

Magnetic-nanoparticle-modified paclitaxel for targeted therapy for prostate cancer.

A nontoxic drug nanocarrier containing carboxyl groups was successfully developed by mixing magnetic nanoparticles (MNPs) of Fe(3)O(4) with the water-soluble polyaniline derivative poly[aniline-co-sodium N-(1-one-butyric acid) aniline] (SPAnNa) and doping with HCl aqueous solution to form SPAnH/MNPs shell/core. SPAnH/MNPs could be used to effectively immobilize the hydrophobic drug paclitaxel (PTX), thus enhancing the drug's thermal stability and water solubility. Up to 302.75 mug of PTX could be immobilized per mg of SPAnH/MNPs. SPAnH/MNPs-bound-PTX (bound-PTX) was more stable than free-PTX at both 25 degrees C and 37 degrees C. Furthermore, bound-PTX was more cytotoxic to human prostate carcinoma cells (PC3 and CWR22R) than free-PTX at 37 degrees C, and the inhibition of cellular growth was even more pronounced when magnetic targeting was applied to the bound-PTX. These data indicate that this magnetically targeted drug delivery system provides more effective treatment of prostate cancer cells using lower therapeutic doses and thus with potentially fewer side-effects.

Age-related reference levels of serum prostate-specific antigen among Taiwanese men without clinical evidence of prostate cancer.

BACKGROUND: To determine the normal distribution of serum prostate-specific antigen (PSA) levels in Taiwanese men without clinical evidence of prostate cancer. METHODS: Between August 2006 and October 2007, healthy Taiwanese men undergoing a routine health examination in our hospital were recruited into the study. All men received a digital rectal examination (DRE) and serum PSA determination and some received a transrectal ultrasound (TRUS). Men with normal DRE findings, and PSA < 4.0 ng/ml, PSA between 4.0 ng/ml and 20 ng/ml with a negative TRUS or abnormal TRUS findings (hypoechoic lesion or others) with a negative biopsy were defined as clinically free of prostate cancer. RESULTS: A total of 7803 participants without clinical evidence of prostate cancer were included in the study. The median PSA value (95th percentile range) was 0.896 ng/ml (3.329) for men 50-59 years old; 1.151 ng/ml (5.114) for men 60-69 years old; 1.623 ng/ml (6.237) for men 70-79 years old and 1.754 ng/ml (6.613) for men older then 80 years. The serum PSA values correlated with age (r = 0.3078, p < 0.001). There were small changes in the median and 95th percentile PSA values in men younger than 50 years old, but large increases in those older than 50 years. CONCLUSIONS: These findings confirm that the serum PSA is directly correlated with age, and offer more efficient PSA reference values for prostate cancer screening tests in Taiwanese men.

The impact of regular biopsy in the first cystoscopic follow-up and other predictors on the recurrence of superficial bladder tumors.

BACKGROUND: To evaluate whether cystoscopic evaluations with bladder biopsy can serve as a significant predictor of superficial bladder tumor recurrence. METHODS: This study examined patients with superficial bladder cancer (Ta/T1). At the first three-month cystoscopic follow-up, subjects were divided into two groups: group 1 received cystoscopic examinations with transurethral bladder biopsy, and group 2 had a cystoscopic examination only. Patients without evidence of recurrence at the first three-month follow-up in these 2 groups (groups 1A and 2A) were compared with respect to recurrence rates and average recurrence time. RESULTS: One hundred and thirty patients were diagnosed with superficial bladder tumors. The mean follow-up time was 22.9 months. The two-year recurrence rates were 33.8% for Ta, 29.8% for T1, 30.4% for low-grade tumors, 31.9% intravesical for high-grade tumors, 26.4% in those treated with bacillus Calmette-Guerin therapy, 33.4% in those who had chemotherapy, 60% in the non-therapy group, 21.8% for single tumors and 47.9% for multiple tumors. Univariate and multivariate analysis revealed that tumor number and intravesical therapy were important predictive factors for tumor recurrence. The average recurrence times in groups 1A and 2A were 18.3 and 12.6 months, respectively, while the respective one-year/two-year recurrence rates were 2.4%/23.8% and 16.9%/22.1%. Univariate analysis showed no statistically significant differences between these two groups. CONCLUSIONS: Our data support tumor number and intravesical therapy as important predictive factors for tumor recurrence. Cystoscopy combined with transurethral bladder biopsy mitigates tumor recurrence in the early years, but not in later years. Therefore, regular cystoscopic evaluation alone is sufficient for longterm patient surveillance.

Mixed germ cell tumor metastatic to the skin: case report and literature review.

BACKGROUND: Testicular cancer is the most common cancer for males aged 15 to approximately 35 years old. The initial presentation is typically an asymptomatic enlarged testicle. The retroperitoneum is the most common metastatic area. Other metastatic sites include the lung, liver, brain, adrenal glands, gastrointestinal tract and spleen. Skin metastasis is a rare event and frequently associated with poor prognosis. CASE PRESENTATION: A 19-year old male was diagnosed testicular mixed germ cell tumor with initial presentation of cutaneous metastasis at scalp and upper abdomen. After radical orchiectomy and four courses of cisplatin-based chemotherapy, the scalp and upper abdominal lesions regressed completely. The size of lung metastases remained unchanged. CONCLUSIONS: For advanced stage testicular cancer, cisplatin-based chemotherapy is still effective to achieve partial response.

Urinary fluorescence in situ hybridization assay for detecting urothelial carcinoma in Taiwanese patients.

OBJECTIVES: To investigate the cytogenetic marker detected by fluorescence in situ hybridization (FISH; UroVysion, Vysis, Inc., Abbott Laboratories, Des Plaines, IL, USA) in the diagnosis of bladder cancer and upper tract urothelial carcinoma (UC) in Taiwanese patients, as FISH has been used in Western countries for detecting UC, but there are limited results in Asian populations. PATIENTS AND METHODS: We analysed polyploidy of chromosome 3, 7, 17 and aneuploidy of chromosome 21, using uroepithelial cells collected at the first void or by instrumental extraction of urine, for bladder cancer, and shedding cells from the upper tract flushed by normal saline via ureteric catheterization or ureterorenoscopy. The criteria for positive tumour cells included three or more positive staining in two or more chromosomes showing polyploidy or <50% staining of the chromosome 9p21. RESULTS: In all, 32 patients with bladder UC and 12 with upper tract UC were assessed. The overall sensitivity for bladder cancer by UroVysion was 96.8%. The sensitivity of the cytology test was 36% for UC of the bladder. The sensitivity for UroVysion in upper tract UC was 12/12 but the specificity was three of nine. CONCLUSIONS: This preliminary report shows that UroVysion was a sensitive screening method for UC of the bladder and upper urinary tract.

Total HLA class I loss in a sarcomatoid renal carcinoma cell line caused by the coexistence of distinct mutations in the two encoding beta2-microglobulin genes.

In renal cell carcinoma (RCC), HLA class I downregulation has been found in about 40% of the lesions examined. Since only scanty information is available about the molecular basis of these defects, we have investigated the mechanism(s) underlying HLA class I antigen downregulation or loss in six RCC cell lines. Five of them express HLA class I antigens although at various levels; on the other hand, HLA class I antigens are not detectable on the remaining cell line, the RCC52 cell line, belonging to a sarcomatoid subtype, even following incubation with IFN-gamma. beta(2)-microglobulin (beta(2) m) was not detected in RCC52 cells. Surprisingly, RCC52 cells harbor two mutations in the beta ( 2 ) m genes in exon 1: a single G deletion (delG) in codon 6, which introduces a premature stop at codon 7, and a CT dinucleotide deletion (delCT), which leads to a premature stop at codon 55. Analysis of eight clonal sublines isolated from the RCC52 cell line showed that the two beta ( 2 ) m gene mutations are carried separately by RCC52 cell subpopulations. The delG/delCT double mutations were detected in two sublines with a fibroblast-like morphology, while the delCT mutation was detected in the remaining six sublines with an epithelial cell morphology. Furthermore, loss of heterozygosity (LOH) of the beta ( 2 ) m gene at STR D15S-209 was found only in the epithelioid subpopulation, indicating loss of one copy of chromosome 15. Immunostaining results of the tumor lesion from which the cell line RCC52 was originated were consistent with the phenotyping/molecular findings of the cultured cells. This is the first example of the coexistence of distinct beta ( 2 ) m defects in two different tumor subpopulations of a RCC, where loss of one copy of chromosome 15 occurs in one of the subpopulations with total HLA class I antigen loss.

Bladder leiomyoma with right hydronephrosis.

The authors describe one female patient with leiomyoma of the bladder who presented with female pseudoprostate and right hydronephrosis. A computed tomography (CT) of the pelvis revealed a soft tissue mass, circumferential and heterogenous in appearance with some calcification inside the mass. Bladder leiomyoma might be one of the differential diagnoses with CT imaging findings of female pseudoprostate.