Huixiang Ji

Celastrol suppresses tumor cell growth through targeting an AR-ERG-NF-κB pathway in TMPRSS2/ERG fusion gene expressing prostate cancer.

The TMPRSS2/ERG (T/E) fusion gene is present in the majority of all prostate cancers (PCa). We have shown previously that NF-kB signaling is highly activated in these T/E fusion expressing cells via phosphorylation of NF-kB p65 Ser536 (p536). We therefore hypothesize that targeting NF-kB signaling may be an efficacious approach for the subgroup of PCas that carry T/E fusions. Celastrol is a well known NF-kB inhibitor, and thus may inhibit T/E fusion expressing PCa cell growth. We therefore evaluated Celastrol’s effects in vitro and in vivo in VCaP cells, which express the T/E fusion gene. VCaP cells were treated with different concentrations of Celastrol and growth inhibition and target expression were evaluated. To test its ability to inhibit growth in vivo, 0.5 mg/kg Celastrol was used to treat mice bearing subcutaneous VCaP xenograft tumors. Our results show Celastrol can significantly inhibit the growth of T/E fusion expressing PCa cells both in vitro and in vivo through targeting three critical signaling pathways: AR, ERG and NF-kB in these cells. When mice received 0.5 mg/kg Celastrol for 4 times/week, significant growth inhibition was seen with no obvious toxicity or significant weight loss. Therefore, Celastrol is a promising candidate drug for T/E fusion expressing PCa. Our findings provide a novel strategy for the targeted therapy which may benefit the more than half of PCa patients who have T/E fusion expressing PCas.

XPC Epigenetic Silence Coupled With p53 Alteration Has a Significant Impact on Bladder Cancer Outcome

PURPOSE Varied XPC genetics are related to bladder cancer susceptibility. We determined whether decreased XPC expression influences bladder cancer malignancy and clinical outcome. MATERIALS AND METHODS Changes in XPC and p53 expression were detected by immunochemistry in 108 bladder cancers, including 29 papillary neoplasms of low malignant potential, and 48 low and 31 high grade lesions, of which 47 were stage Ta-T1 and 61 were stage T2-T3. XPC mRNA and methylation were evaluated in fresh tissue by real-time reverse transcriptase and methylation specific polymerase chain reaction. The clinical value of altered XPC and p53 expression was analyzed in 66 bladder cancers, including 6 papillary neoplasms of low malignant potential, and 41 low and 19 high stage lesions, of which 26 were stage Ta-T1 and 40 were stage T2-T3, by the Kaplan-Meier method and Cox proportional hazards regression. RESULTS The XPC defect was associated with bladder cancer higher pathological grade, metastasis and p53 mutation. Patients with XPC(-)/p53(+) had shorter survival than those with bladder cancer without XPC(-)/p53(+) (p = 0.0127). Cox regression analysis showed that XPC expression may be a potential predictive factor for bladder cancer (p = 0.043). In bladder cancer xpc gene hypermethylation was significantly higher than in normal mucosa (p = 0.0437). CONCLUSIONS Lower mRNA may be the result of XPC hypermethylation in bladder cancer. Epigenetic defects in the XPC gene impact bladder cancer malignant behavior and may also predict poor outcome in some bladder cancer cases, as characterized by p53 pathway alteration.

Better Compliance Contributes to Better Nocturnal Continence With Orthotopic Ileal Neobladder Than Ileocolonic Neobladder After Radical Cystectomy for Bladder Cancer

OBJECTIVES To investigate, in a randomized controlled study, the degree of continence after the creation of orthotopic ileocolonic and ileal neobladders after cystectomy and to explore a possible mechanism for the difference in continence between these 2 types of orthotopic neobladder. METHODS From 2003 to 2007, 71 male patients underwent orthotopic lower urinary tract reconstruction with either an ileocolonic or ileal neobladder after radical cystectomy. The degrees of continence and voiding patterns were individually evaluated using urodynamic examinations and a detailed patient questionnaire. The abnormal upper tract was evaluated using intravenous urography and ultrasonography. RESULTS Complete daytime continence was achieved in 90.9% and 89.4% of the patients and functional nocturnal continence 48.5% and 76.3% of patients in the ileocolonic neobladder and ileal neobladder groups, respectively. The urodynamic data showed that the initial volume of both the ileocolonic and the ileal neobladder appeared to not be significantly different statistically, although the compliance of the ileocolonic neobladder was lower than that of the ileal neobladder (P < .05). No difference was found in the parameters such as flow rate, urethral profile length, maximal urethral pressure, or neobladder neck pressure between the 2 neobladder types. CONCLUSIONS Although the ileocolonic and ileal neobladders can both achieve a large initial volume, the ileal neobladder has an advantage in the aspect of obtaining satisfactory nocturnal continence because of its greater compliance compared with that of the ileocolonic neobladder.

Cathepsin L is Associated with Proliferation and Clinical Outcome of Urothelial Carcinoma of the Bladder

Increasing evidence suggests that the activity of cysteine proteases, including cathepsin L, is important in cancer cell invasion and metastasis. This study was designed to investigate the clinicopathological and prognostic significance of cathepsin L in human urothelial carcinoma of the bladder (UCB). Standard immunohistochemistry was used to determine the presence of cathepsin L and Ki-67 (a marker of proliferation) in paraffin-embedded specimens of 82 human UCB cases. Cathepsin L protein was localized in the cytoplasm of the malignant UCB cells, and was significantly associated with the pathological tumour stage (invasiveness), tumour grade, survival, local tumour recurrence during follow-up, the occurrence of distant metastasis during follow-up and the presence of Ki-67 protein. Multivariate analysis using the Cox proportional hazards model revealed that cathepsin L immunopositivity and pathological tumour stage (invasiveness) were independent significant prognostic factors for overall survival. This study showed that cathepsin L provides significant prognostic information and that it might be a useful therapeutic target in the future.

Identification and Management of Emptying Failure in Male Patients With Orthotopic Neobladders After Radical Cystectomy for Bladder Cancer

OBJECTIVE To treat neobladder emptying failure after radical cystectomy in patients with bladder cancer. The etiology of neobladder emptying failure should be identified. METHODS We analyzed the outcome of neobladder emptying in 231 male patients who received neobladder reconstruction after radical cystectomy. The clinical characteristics, urodynamic evaluation, and treatment information were collected from all patients with emptying failure. RESULT The total occurrence of neobladder emptying failure was 37 of 231 (16%). Emptying failure was a result of mechanical obstruction in 25 (10.8%) patients; obstructions were caused by strictures of the neobladder-urethral anastomosis (13 cases, 5.6%), anterior urethral strictures (3 cases, 1.2%), obstructive mucosal valves (2 cases, 0.9%), primary cystolithiasis (1 case, 0.4%), mucus plugs (2 cases, 0.9%), urethral tumor recurrence (2 cases, 0.9%), and pelvic tumor recurrence (2 cases, 0.9%). In 21 of 25 patients with mechanical obstructions, bladder function was completely recovered via an endourological approach. However, in 12 of patients with dysfunctional voiding, 3 patients presented higher compliance of neobladder. Two patients were found with a narrower posterior urethral angle. Eventually, 10 patients of 12 with dysfunctional voiding performed intermittent self-catheterization. CONCLUSIONS The obstructive outlet was the primary cause of emptying failure in neobladders. Most of the patients with mechanical obstructions could obtain satisfactory neobladder emptying by a minimally invasive surgical approach. However, nearly all the patients with dysfunctional voiding will have to receive clean intermittent catheterization until the mechanisms causing failure are better understood.

Long-Term Evaluation of Patients Undergoing Genitoplasty due to Disorders of Sex Development: Results from a 14-Year Follow-Up

Purpose. To summarize the experience in treating patients with genitoplasty due to disorders of sex development in China. Methods. The operative procedures, gender of rearing, surgical outcome, and psychosocial and family adjustments of 262 patients were reviewed retrospectively. Results. At initial diagnosis, the mean age was 14.3 ± 2.8 years (range: 2–38 years). There were 96 children, 133 adolescents, and 33 adults. Follow-up was done every 6 months. Patients with female sex assignment had no urinary incontinence or voiding difficulty. Five patients underwent the second surgery (3%); vaginal dilation was performed in 35 patients with postoperative vaginal stenosis; 12 patients (7.4%) were unsatisfactory with the outcome. For patients with male sex assignment, the median length of penis was 2.2 cm in prepubertal patients, 4.2 cm in pubertal patients, and 5.0 cm in adults; 39 patients developed postvoid dribbling (39%); 21 patients underwent a second surgery (21%); urethral dilation was done in 28 patients (28%) due to urethral stricture; 38 patients were unsatisfactory with the outcome (38%). In addition, 136 patients (83%) with female sex assignment and 54 (54%) with male sex assignment had favorable psychosocial adjustment. Conclusions. Patients with male sex assignment have more surgical complications and difficulties in psychosocial adjustment as compared to those with female sex assignment.

Downregulated XPA promotes carcinogenesis of bladder cancer via impairment of DNA repair

Bladder cancer is the most common malignant tumor of urinary system, largely resulting from failure of repair of DNA damage to the environmental insults. The function of XPA in nucleotide excision repair pathway has been well documented. However, participation of XPA in the repair of DNA double-strand break remains unknown. Here, we reported that bladder cancer expressed low XPA levels compared to adjacent non-tumor bladder tissue, and this phenotype was closely associated with chromosomal aberrations. Moreover, downregulated XPA appeared to increase incidence of chromosome aberration. XPA reduction increased cell viability of a bladder cancer cell line RT4, while XPA re-expression decreased the cell viability of RT4 cells. Since high mutation frequency is the basis of mutations of oncogenes and anti-oncogenes, and may be the essence of bladder cancer susceptibility, our study suggests that downregulated XPA may promote carcinogenesis of bladder cancer via impairment of DNA repair.

Use of urostomy bags in the management of perioperative urine leakage after radical cystectomy.

Background: Urine leakage is a common complication in patients with bladder cancer after radical cystectomy and neobladder reconstruction. Objective: The aim of this study was to evaluate the clinical value of the use of urostomy bags in the management of urine leakage in patients with bladder cancer after radical cystectomy. Methods: Urine leakage during the perioperative period was retrospectively analyzed in 483 patients with bladder cancer who underwent radical cystectomy from 2004 to 2010. Before 2008, all patients with urine leakages were treated by routine dressing changes (group A). After 2008, the leakages were managed with urostomy bags (group B). The perioperative quality of life (EQ-5D) and cost for urine leakage for both groups were compared in this controlled study. Results: The average cost in management of preoperative urine leakage was significantly higher in group A than in group B as well as the patients with extravasations of urine or lymphoceles. Patients in group B had an overall better perioperative life quality compared with group A. In particular, the score for pain/discomfort was significantly higher in group A than in group B. Conclusions: The management of perioperative urine leakage with urostomy bags avoided constant body wetness and significantly increased the quality of life and reduced the special costs of urine leakage in patients with bladder cancer after cystectomy. Implications for Practice: Early use of urostomy bag is a good choice for perioperative urine leakage in patients with bladder cancer after radical cystectomy and neobladder reconstruction.

Ultrastructural features and possible functional role of kit-positive interstitial cells in the guinea pig corpus cavernosum.

The objective of the present study was to identify kit-positive interstitial cells (ICs) in guinea pig corpus cavernosum and examine their relationships with adjacent smooth muscle cells (SMCs) and intramural nerves. In addition, we investigated the possible involvement of ICs in nitric oxide (NO)-mediated relaxation of corpus cavernosum smooth muscle (CCSM). ICs were identified by their immunoreactivity to the kit receptor, a cell surface marker encoded by c-kit proto-oncogene and specific for interstitial cells of Cajal. ICs were abundantly distributed in guinea pig corporal tissues. Ultrastructural investigation by conventional transmission electron microscopy revealed the ultrastructural features of ICs and gap junctions located between ICs and adjacent SMCs, furthermore, a close contact between ICs and intramural nerves for the first time. Western blot analysis of purified ICs by fluorescence-activated cell sorting revealed coexpression of soluble guanylate cyclase (sGC)α1, sGCβ1 and kit receptor tyrosine kinase protein in them. These observations imply that ICs express the NO-sensitive sGC molecule and may be involved in the NO-mediated relaxation of CCSM in the guinea pig corpus cavernosum.

Human Telomerase Reverse Transcriptase Transfection Reduces Apoptosis in Human Penile Smooth Muscle Cells and Slows Down Cellular Aging

INTRODUCTION Erectile dysfunction (ED) is one of the most common diseases in male urology that greatly affects the quality of life in senior people. Relaxation of corpus cavernosum smooth muscle is the key to penile erection. AIM To explore effects of human telomerase reverse transcriptase (hTERT) gene transfection on biological behaviors of human penile smooth muscle cells. METHODS Human penile smooth muscle cells were grown in primary culture. A fluorescent eukaryotic expression vector, hTERT-internal ribosome entry site 2 (IRES2)-enhanced green fluorescent protein (EGFP), was constructed and transfected into human penile smooth muscle cells using Lipofectin reagent. MAIN OUTCOME MEASURE The telomerase activity, mitotic index, cell apoptosis, and cell growth curves of transfected smooth muscle cells were determined; the potential formation of malignant phenotypes in these transfected cells was investigated. RESULTS Telomerase activity, mitotic index, and cell growth of hTERT-transfected cells were significantly higher than those of nontransfected cells and cells transfected with the empty EGFP vector, while apoptosis rates were the lowest in hTERT-transfected cells. No changes in cell morphology, chromosome number, and tumorigenicity were observed between hTERT-transfected cells and control cells. CONCLUSION In this study, for the first time, the hTERT gene was transfected into human penile smooth muscle cells, and the gene increased telomerase activity in cells, reduced cell apoptosis, and slowed down cell aging. We believe that this finding is of potential clinical value in the prevention and treatment of organic ED.