DNA hypermethylation in papillary renal cell carcinoma

What’s known on the subject? and What does the study add? Epigenetic alterations play an essential role during carcinogenesis, and earlier studies suggested that global histone modification levels are predictive for patients’ outcome in various tumour entities (e.g. prostate, lung, breast and kidney cancer). So far, only smaller studies on histone H3K9 acetylation in patients with papillary urothelial neoplasm of low malignant potential (PUNLMP) and histone H2AX phosphorylation in non‐muscle invasive bladder cancer exist; both studies indicate prognostic relevance for bladder cancer. We demonstrate that histone methylation levels (H3K4, H4K20) decrease from normal tissue over non‐muscle invasive and muscle invasive bladder cancer to bladder cancer metastases. Histone modifications are correlated to advanced pathological stage in non‐muscle and muscle invasive bladder cancer. Furthermore, H4K20me3 appeared to be predictive for bladder cancer specific survival in patients with muscle‐invasive bladder cancer undergoing radical cystectomy.

OBJECTIVE

To investigate the pattern of DNA CpG island hypermethylation in papillary renal cell carcinoma (pRCC).

MATERIAL AND METHODS

DNA from pRCC (n= 32) and adjacent normal tissue (n= 15) was isolated. A quantitative methylation‐specific PCR was performed to analyse the methylation pattern at APC (actin beta), CDH1 (E‐cadherin), GSTP1 (glutathione S‐transferase pi 1), RASSF1A (Ras association domain family member 1A) and TIMP3 (TIMP metallopeptidase inhibitor 3); a sequence of ACTB without CpG was used to normalize for DNA input and to calculate the relative amount of methylated DNA (normalized index of methylation, NIM).

RESULTS

RASSF1A hypermethylation was observed in most pRCC and normal samples (100 vs 94.4%), but the median NIM was significantly higher in pRCC samples (2.11 vs 0.61; P < 0.001). RASSF1A hypermethylation allowed discrimination of pRCC and normal tissue with a sensitivity of 87.5% and a specificity of 73.3% as determined via receiver operator characteristic analysis (area under curve = 0.814). Hypermethylation at APC (3.0 vs 6.7%), CDH1 (15.6 vs 0%), GSTP1 (21.9 vs 6.7%) and TIMP3 (6.3 vs 0%) was infrequent in pRCC and normal tissue. CDH1 was significantly correlated with pathological stage (P= 0.015), and patients with methylated CDH1 methylation showed a trend towards shorter recurrence‐free survival (log‐rank P= 0.057). The number of methylated gene sites was correlated with pathological stage (P= 0.007) and lymph node metastasis (P= 0.008).

CONCLUSIONS

DNA hypermethylation at RASSF1A is common in pRCC tissue irrespective of the histological subtype, but also frequently seen at lower levels in normal adjacent tissue. Aberrant hypermethylation could be a prognostic marker for pRCC.     

Hypermethylation of E-cadherin,p16, p14, and RASSF1A genes in pathologically normal urothelium predict bladder recurrence of bladder cancer after transurethral resection

ObjectivesThis study investigated the hypermethylation of E-cadherin, p16, p14, and RASSF1A in pathologically normal urothelium to predict recurrence of bladder cancer after transurethral resection.Materials and methodsSamples of bladder tumor and paired pathologically normal urothelium were obtained from 50 bladder cancer patients. The status of promoter hypermethylation in these four genes was investigated by methylation-specific polymerase chain reaction. The clinicopathologic data in these patients were also analyzed in order to evaluate the clinical implication of aberrant methylation in bladder cancer recurrence.ResultsHypermethylation of E-cadherin (30%), p16 (16%), p14 (14%), and RASSF1A (36%) was detected in the pathologically normal urothelium samples. Promoter hypermethylation occurred frequently in both pathologically normal urothelium and tumor samples from bladder cancer patients, and increased with progression from normal to bladder cancer at E-cadherin (P = 0.067), p16 (P < 0.001), p14 (P = 0.01), and RASSF1A (P = 0.01). No significant correlation was observed between hypermethylation in any genes and muscle/organ invasion and stage/grade, except p14. However, p14 hypermethylation in pathologically normal urothelium samples was associated with shorter recurrence-free interval (P = 0.019).Conclusionsp14 hypermethylation could be involved in early stage of bladder carcinogenesis, and p14 hypermethylation in pathologically normal urothelium samples should be considered a predictor of bladder cancer recurrence.     

Association of RASSF1A genotype and haplotype with the progression of clear cell renal cell carcinoma in Japanese patients

What's known on the subject? and What does the study add? Ras association domain family 1A (RASSF1A) is a tumour suppressor and regulates cell cycle, apoptosis and microtubule stability. This is the first study to identify associations between RASSF1A polymorphisms and clinicopathological parameters and survival in patients with clear cell renal cell carcinoma (CCRCC). RASSF1A genotyping may be useful for predicting the prognosis of the clinical course of CCRCC, and this finding might provide a better understanding of the mechanism underlying the development and progression of CCRCC. However, functional and prospective studies with a larger number of patients are needed to confirm the results.

OBJECTIVE

• ? To compare Ras association domain family 1A (RASSF1A) genotypes or haplotypes with clinicopathological characteristics and survival rates of patients with clear cell renal cell carcinoma (CCRCC).

PATIENTS AND METHODS

• ? The study cohort comprised 224 Japanese patients who underwent radical nephrectomy and had CCRCC confirmed by histopathological analysis.
• ? Three common polymorphisms in the RASSF1A gene, 133Ala/Ser (G/T), ‐710C/T and ‐392C/T, were genotyped using TaqMan assays and haplotypes were analysed using appropriate software.

RESULTS

• ? Patients with CCRCC with RASSF1A ‐710TT genotype exhibited a significantly higher tumour stage and higher stage grouping than those with ‐710CC or ‐710CT (P = 0.005 and P = 0.032, respectively).
• ? There was no significant association between 133Ala/Ser or ‐392C/T genotype and clinicopathological characteristics.
• ? RASSF1A 133Ala‐710T‐392T haplotype and ‐710TT genotype were significantly associated with poorer recurrence‐free survival rates (P = 0.038 and P = 0.007, respectively).

CONCLUSIONS

• ? This is the first study to identify associations between RASSF1A polymorphisms and clinicopathological parameters and survival in patients with CCRCC.
• ? RASSF1A genotyping may be useful in predicting the prognosis of the clinical course of CCRCC, and this finding might provide a better understanding of the mechanism underlying the development and progression of CCRCC.
• ? Functional and prospective studies with a larger number of patients are needed to confirm the results.

     

Prognostic value of syndecan-1 expression in patients treated with radical prostatectomy

OBJECTIVE

To investigate the association of syndecan‐1 expression with pathological features and disease progression in patients treated with radical prostatectomy (RP) as syndecan‐1 plays a role in the regulation of cell proliferation, migration, and differentiation and its expression is altered in various malignancies.

PATIENTS AND METHODS

Syndecan‐1 immunostaining was performed on a tissue microarray containing cores from 232 consecutive patients treated with RP and bilateral lymphadenectomy for clinically localized prostatic adenocarcinoma. Patients were categorized as having features of aggressive progression if they had evidence of metastases, an after progression prostate‐specific antigen (PSA) doubling time of <10 months, and/or failure to respond to local salvage radiation therapy. Expression was defined as ≥ 10% cells staining for syndecan‐1.

RESULTS

Syndecan‐1 was expressed in 86 patients (37.1%). Expression of syndecan‐1 was associated with higher PSA levels (P = 0.004), higher pathological Gleason sum (P = 0.027) and lymph nodes metastases (P = 0.027). Patients with syndecan‐1 expression were at significantly greater risk of PSA‐progression after surgery (P = 0.034) in univariate but not in multivariate analysis. Patients with features of aggressive progression (n = 22) were more likely to express syndecan‐1 than those with features of nonaggressive progression (63.6% vs 36.4%, P = 0.010). Patients with syndecan‐1 expression were at significantly greater risk of aggressive progression after surgery (P = 0.005) in univariate but not in multivariate analysis.

CONCLUSIONS

Expression of syndecan‐1 was associated with established features of biologically aggressive prostate cancer and PSA‐progression in univariate analysis. These findings suggest a role for syndecan‐1 in prostate carcinogenesis and progression.     

Improved cancer specific-survival in patients with carcinoma invading bladder muscle expressing cyclo-oxygenase-2

Study Type – Prognosis (case series) Level of Evidence 4

OBJECTIVE

To determine whether the expression of cyclo‐oxygenase (COX)‐2 has an influence on survival and on the response to chemotherapy in invasive bladder cancer.

PATIENTS AND METHODS

A population of 266 patients from a tertiary university centre with carcinoma invading bladder muscle without evidence of metastasis at time of cystectomy was analyzed retrospectively. COX‐2 expression was evaluated immunohistochemically with a monoclonal anti‐COX‐2 antibody. All pertinent clinical and pathological parameters were reviewed and correlated with risk factors influencing outcome, including disease‐specific and overall survival, as well as COX‐2 expression. Immunoreactivity was categorized as positive if COX‐2 staining was present in >5% tumour cells.

RESULTS

The expression of COX‐2 was not influenced by tumour stage, grade or nodal status, nor any other parameters. The risk factors that influenced disease‐specific survival in carcinoma invading bladder muscle on multivariate analysis were lymph node status (hazards ratio, HR = 2.46 for N1, P= 0.001, HR = 2.90 for N2, P < 0.001, HR = 5.19 for N3, P= 0.012), use of neoadjuvant chemotherapy (HR = 3.54; P= 0.004) or adjuvant chemotherapy (HR = 0.57, P= 0.014) and COX‐2 expression (HR = 0.64 if >5% cells had positive expression; P= 0.025). Kaplan–Meier analysis showed an increased disease‐specific survival (P= 0.0063), as well as longer recurrence‐free survival (P= 0.003), in patients with muscle‐invasive bladder tumours expressing COX‐2 in >5% of the cells. A tendency was also observed in a subgroup with positive nodes treated with adjuvant chemotherapy (P= 0.093).

CONCLUSIONS

The overexpression of COX‐2 is associated with a better recurrence‐free and disease‐specific survival in a large cohort of 266 patients with carcinoma invading bladder muscle treated by cystectomy. A trend for increased disease‐specific survival was also observed for patients with COX‐2 overexpression and positive nodes who received adjuvant chemotherapy. Potential of COX‐2 as a prognostic marker in bladder cancer should be considered.     

A double‐blind placebo‐controlled study of the efficacy and safety of pentoxifylline in early chronic Peyronie’s disease

Study Type – Therapy (RCT)
Level of Evidence 1b

OBJECTIVE

To analyse the safety and efficacy of pentoxifylline sustained‐release (PTX‐SR) treatment in patients with early chronic Peyronie’s disease (PD).

PATIENTS AND METHODS

In all, 228 patients with a mean (sd ) age of 51 (9) years who had early chronic PD were randomized to receive 400 mg PTX‐SR (Apo‐Pentoxifylline, Apotex Inc., Toronto, Canada) twice daily (group 1, 114) or similar regimen of placebo (group 2, 114) for 6 months. A medical history was taken and the men had a complete physical examination. The following variables were assessed before and after therapy: penile curvature and penile artery spectral traces (end‐diastolic velocity, EDV, peak systolic velocity, PSV, and resistivity index, RI, of the right and left cavernous arteries assessed with dynamic penile duplex ultrasonography), plaque characteristics (assessed by penile X‐ray and penile ultrasonography), pain (assessed by visual analogue scale), erectile function (assessed by the International Index of Erectile Function, IIEF questionnaire), treatment satisfaction (assessed by Erectile Dysfunction Inventory of Treatment Satisfaction questionnaire), and side‐effects. Patient perception of penile curvature and plaque size, and mean weekly intercourse attempts were also assessed.

RESULTS

Overall, 36.9% of patients who received PTX‐SR reported a positive response, vs only 4.5% in the placebo group. Of patients in PTX‐SR group, 12 (11%) had disease progression, vs 46 (42%) in placebo group (P = 0.01). Improvement in penile curvature (P = 0.01), and plaque volume (P = 0.001) was significantly greater in patients treated with PTX‐SR than placebo. The increase in IIEF total score was significantly higher in the PTX‐SR group (P = 0.02). Mean PSV changes after therapy compared to baseline were statistically significant between PTX‐SR (right, +11.4%, left, +11.7%) and placebo‐treated (+0.2% and ?4.2%, respectively) patients (both P = 0.04).

CONCLUSIONS

PTX‐R was moderately effective in reducing penile curvature and plaque volume in patients with early chronic PD. Further studies with different treatment regimens are needed to better elucidate the beneficial effects of PTX‐SR in PD.     

The prognostic value of peritumoral regulatory T cells and its correlation with intratumoral cyclooxygenase‐2 expression in clear cell renal cell carcinoma

OBJECTIVE

To investigate the prognostic value of regulatory T cells (Tregs) and its correlation with cyclooxygenase‐2 (COX‐2) expression in clear cell renal cell carcinoma (RCC).

PATIENTS AND METHODS

CD4+, Foxp3+ tumour‐infiltrating lymphocytes and tumour COX‐2 expression were assessed by immunohistochemistry in tissue microarrays containing RCC from 125 patients. Prognostic effects of low and high expression were evaluated by Cox regression and Kaplan–Meier analysis using the median values as thresholds. The expression of Tregs and COX‐2 were compared with the clinicopathological variables. In addition, Tregs and its correlation with COX‐2 expression was also analysed.

RESULTS

Peritumoral Tregs were positively correlated with intratumoral COX‐2 expression (Spearman rank correlation 0.336, P < 0.001). Peritumoral Tregs were associated with TNM stage (P = 0.001) and tumour size (P = 0.002), while intratumoral COX‐2 expression was associated with TNM stage (P = 0.018) and grade (P = 0.013). Using multivariate analysis, increased peritumoral Tregs, higher TNM stage (III + IV), larger tumour size (≥7 cm) and higher nuclear grade (III + IV) were independent predictors for significantly shorter overall survival and disease‐free survival.

CONCLUSIONS

Increased peritumoral Tregs are associated with worse prognosis in clear cell RCC. The high intratumoral COX‐2 expression may be the underlying reason for the aberrant gathering of Tregs. These results suggest that clinical application of COX‐2 inhibitors may benefit those patients with higher intratumoral COX‐2 immunostaining by reducing the transformation of Tregs in RCC.     

Impact of PD‐L1 expression and human papillomavirus status in anti‐PD1/PDL1 immunotherapy for head and neck squamous cell carcinoma—Systematic review and meta‐analysis

Programmed cell death‐1 (PD‐1) pathway inhibition in head and neck squamous cell carcinoma (HNSCC) has demonstrated inconsistent efficacy regarding human papillomavirus (HPV) status and PD‐L1 expression. This study compared outcomes in HNSCC in the context of PD‐L1 and HPV expression. Outcomes: PD‐L1 and HPV expression; overall survival (OS), and tumor response (ORR). 1088 patients received PD‐1/L1 inhibitors. Four methodologies were identified in determining PD‐L1 expression, most commonly using the Dako PD‐L1 IHC 22C3 pharmaDx assay. Using a 1% threshold, ORR was greater for PD‐L1 expressers vs non‐expressers (18.9%, CI 16.1‐21.8 v 8.8% CI 5.3‐13.7, P = 0.009), as was OS at 6 months (60.6%, CI 49.2‐71.4 v 49.0%, CI 39.1‐59.0, P = 0.04) but not at 12 or 18 months. No advantages were identified for HPV expressers. Patients expressing PD‐L1 may have a better tumor response and OS. No impact on survival or response was observed based on HPV status.     

Exposed proliferation antigen 210 (XPA-210) in renal cell carcinoma (RCC) and oncocytoma: clinical utility and biological implications

What's known on the subject? and What does the study add? The exposed proliferation antigen 210 (XPA‐210) of the proliferation marker thymidine kinase 1 (TK1) showed higher expression levels in metastatic renal cell carcinoma. The current study used a new XPA‐210 antibody to clarify the role of TK1 tissue expression in the largest reported cohort of different renal cell carcinoma types and oncocytomas.

OBJECTIVE

• ? To determine the clinical role of the exposed proliferation antigen 210 (XPA‐210) of the proliferation marker thymidine kinase 1 (TK1) in a large cohort of different renal cell carcinoma (RCC) types, oncocytomas and normal renal tissues samples, as TK1 is reported to be of clinical significance in several cancer entities and is suggested as a prognostic serum biomarker for RCC.

PATIENTS AND METHODS

• ? Expressions of XPA‐210 were determined immunohistochemically in 40 clear cell RCCs (ccRCC), 25 papillary RCCs (papRCC), 17 chromophobe RCC (chRCC), 27 oncocytomas and 64 normal renal parenchyma paraffin‐embedded specimens.
• ? Immunohistochemistry was performed with a monoclonal anti‐XPA‐210 antibody. Staining was measured by the percentage of positive cells.
• ? Expression was compared between subgroups and correlated with respective clinical data using one‐way analysis of variance with post hoc Tukey‐Kramer analyses.

RESULTS

• ? XPA‐210 staining in the RCC subgroup was significantly different from the oncocytomas (mean [sem ] 4.1 [0.4] vs 2.2 [0.4]; P = 0.004) and from normal renal tissue (1.0 [0.1]; P < 0.001], whereas oncocytomas did not differ from normal renal parenchyma staining (P = 0.18).
• ? Subdivided into RCC groups, only ccRCC (mean [sem ] 5.1 [0.6]; P < 0.001) and papRCC (4.4 [0.6]; P < 0.001) varied from normal renal parenchyma, whereas chRCC (1.4 [0.3]; P = 0.99) did not.
• ? RCC XPA‐210 staining was significantly associated with higher tumour stage (T = 3, P = 0.002) and grade (G = 3, P = 0.001).

CONCLUSIONS

• ? The malignant character of RCC is reflected by higher XPA‐210 expression as compared with oncocytomas and normal kidney.
• ? The ccRCC and papRCC subgroups had higher XPA‐210 levels.
• ? XPA‐210 could be considered a potential marker for the assessment of the proliferative activity in primary RCC.

     

β‐catenin expression and claudin expression pattern as prognostic factors of prostatic cancer progression

OBJECTIVE

To investigate the patterns of expression of the junctional proteins β‐catenin and claudins in different prognostic groups of patients with prostatic cancer, to determine their value as prognostic markers.

PATIENTS AND METHODS

We evaluated the samples of 30 patients who had a radical prostatectomy for organ‐confined cancer (pT2N0M0), men with clinically advanced cancer, and a control group with benign prostatic hyperplasia. Using immunohistochemistry applied to tissue microarrays, each group was evaluated for claudin‐1, ‐2, ‐3, ‐4, ‐5, ‐7, ‐8 and ‐10, and β‐catenin expression.

RESULTS

There were differences among the three groups in the expression of claudin‐1 (P = 0.001), ‐2 (P = 0.014), ‐3 (P = 0.027), ‐4 (P = 0.001), ‐8 (P = 0.001) and β‐catenin (P = 0.002), regardless of Gleason score. By contrast, claudin‐5, ‐7 and ‐10 patterns were not significantly different among the groups. Furthermore, claudin‐1 (P = 0.014) and ‐4 (P = 0.004) could be used to distinguish between those patients who had metastases and those who did not.

CONCLUSIONS

The pattern of claudin expression could be a novel diagnostic marker in re‐classifying adenocarcinomas, and an additional sensitive predictive factor for a clinically poor prognosis. Our results suggest that patients with organ‐confined and advanced cancer are subsets with distinct claudin expression profiles, and that claudin‐4 is related to cellular differentiation in prostate cancer, which is not only the receptor molecule for the Clostridium perfringens enterotoxin, and thus a theoretical future therapeutic target for prostate cancer, but also a marker of progression.     

P-cadherin as a prognostic indicator and a modulator of migratory behaviour in bladder carcinoma cells

OBJECTIVE

To identify changes associated with P‐cadherin expression in bladder cancer and evaluate the potential role of such events in determining the clinical outcome and cell behaviour, as the function of P‐cadherin in normal epithelium is unknown, as is its potential role in neoplastic progression in different cancers.

MATERIALS AND METHODS

In all, 536 bladder tumour specimens from 408 patients were assembled in seven tissue microarrays. Paraffin sections from each array were processed for immunohistochemistry to assess the expression of P‐cadherin. The expression of P‐cadherin was forced using lipofectin, followed by an assessment of migration and invasion potential using standard in vitro assays.

RESULTS

The absence of P‐cadherin staining was associated with muscle‐invasive disease, grade 3 (P < 0.001) and nodal disease (P = 0.009). Similar results were obtained when considering cytoplasmic and unrestricted localization of P‐cadherin (P < 0.001), except for nodal involvement. The group with cytoplasmic location of P‐cadherin showed a shorter cancer‐specific survival than the group with membrane location of P‐cadherin (P = 0.03). Forced expression of P‐cadherin in EJ and UM‐UC‐3 cells, that constitutively lack P‐cadherin expression, resulted in modulation of catenin expression and enhanced migration of EJ and UM‐UC‐3/P‐cadherin transfectants (>200%).

CONCLUSIONS

These results showed that loss of expression, cytoplasmic relocation or unrestricted tissue location of P‐cadherin was associated with a poor clinical outcome and prognosis in bladder cancer. From the in vitro work it is evident that P‐cadherin plays a role in regulating the migration potential of bladder carcinoma cells.     

Interest of methylated genes as biomarkers in urothelial cell carcinomas of the urinary tract

Urothelial cell carcinomas (UCC), including bladder cancer and upper urinary tract cancer, are the second most common malignancy of the urogenital tract after prostate cancer. It is a critical issue to differentiate accurately those patients whose tumour will recur and/or progress after initial treatment from those without recurrence and/or progression because tumours with a similar morphology behave differently. Patients undergo lifelong cystoscopic and cytology surveillance to detect subsequent tumour recurrence. New noninvasive methods for the diagnosis and surveillance of UCCs are required. DNA methylation is an important epigenetic mechanism of gene regulation and plays essential roles in tumour initiation and progression. Currently, aberrant promoter hypermethylation has been investigated in specific genes, i.e. tumour‐suppressor genes, proto‐oncogenes, genes involved in cell adhesion, and genes of cell cycle regulation. E‐cadherin has been shown to be an independent marker of prognosis. Other genes, e.g. APC, RASSF1a, TNFRSF25, EDNRB, and p14, are implicated in tumour progression. IGFBP3 and APAF‐1 are independent markers of recurrence. APAF‐1 is also correlated with tumour stage and grade. In urine, hypermethylation of DAPK, RARβ, E‐cadherin and p16 has been shown to have a good sensitivity and specificity for bladder cancer detection. Several studies found that analysis of hypermethylation using a panel of tumour‐suppressor genes yielded superior results to cytology in the detection of bladder cancer and its progression. Hence, the different panels (e.g. RASSF1a/APC/p14, RAR_/DAPK/E‐cadh/p16, p16/p14/MGMT/GSTP1, and RASSF1a/E‐cadh/APC) are of interest in the detection of bladder cancer. The last panel and RASSF1a/E‐cadh/APC/TNFRSF25/EDNRB are also interesting for tumour progression. There is evidence that the extent of the mutator and methylator phenotypes in UCCs differs with tumour location, perhaps suggesting that carcinogens affect the urinary tract in different ways. For all loci studied except DAPK, there was more frequent methylation in UUT‐UCCs than in the bladder cancers; this difference was statistically significant for hMLH1, RARB, E‐cadherin, p16 and MINT31. In contrast to UUT‐UCCs, hMLH1 and MINT31 were rarely methylated in bladder tumours, suggesting that they play a role in UUT carcinogenesis but not bladder cancer.     

Uridine 5'diphospho‐glucuronosyltransferase 1A expression as an independent prognosticator in urothelial carcinoma of the upper urinary tract

Objective

To determine the expression status of uridine 5'diphospho‐glucuronosyltransferase 1A, a major phase II drug metabolism enzyme, in upper urinary tract urothelial carcinoma, as well as to assess its prognostic significance.

Methods

We immunohistochemically stained for uridine 5'diphospho‐glucuronosyltransferase 1A in tissue microarray consisting of 99 upper urinary tract urothelial carcinoma samples and paired non‐neoplastic urothelial tissues. We also assessed the effect of uridine 5'diphospho‐glucuronosyltransferase 1A knockdown on urothelial cancer cell growth.

Results

Uridine 5'diphospho‐glucuronosyltransferase 1A was positive in 92.9% (27.3% weak [1+], 37.4% moderate [2+], 28.3% strong [3+]) of tumors, which was significantly (P < 0.001) lower than in benign urothelial tissues (98.8%; 3.5% 1+, 18.8% 2+, 76.4% 3+). All 37 (100%) non‐muscle‐invasive versus 55 (88.7%) of 62 muscle‐invasive tumors (P = 0.043) were immunoreactive for uridine 5'diphospho‐glucuronosyltransferase 1A. The rates of moderate‐to‐strong uridine 5'diphospho‐glucuronosyltransferase 1A expression and its positivity were also strongly associated with the absence of concomitant carcinoma in situ (P = 0.034) and lymphovascular invasion (P = 0.016), respectively. However, there were no statistically significant associations between uridine 5'diphospho‐glucuronosyltransferase 1A expression and tumor grade or pN/M status. Uridine 5'diphospho‐glucuronosyltransferase 1A loss in M0 tumors was strongly associated with lower progression‐free survival (P < 0.001) and cancer‐specific survival (P < 0.001) rates. Multivariate analysis further identified a strong correlation of uridine 5'diphospho‐glucuronosyltransferase 1A positivity with reduced cancer‐specific mortality (hazard ratio 0.28, P = 0.018). Meanwhile, uridine 5'diphospho‐glucuronosyltransferase 1A knockdown in urothelial cancer cells resulted in significant increases in their viability and migration.

Conclusions

These results suggest a preventive role of uridine 5'diphospho‐glucuronosyltransferase 1A signals in the development and progression of upper urinary tract urothelial carcinoma. Loss of uridine 5'diphospho‐glucuronosyltransferase 1A expression might serve as an independent predictor of poor prognosis in patients with upper urinary tract urothelial carcinoma.     

Self‐ratings of genital anatomy,sexual sensitivity and function in men using the ‘Self‐Assessment of Genital Anatomy and Sexual Function,Male’ questionnaire

OBJECTIVE

To assess the perceptions of healthy men of their genital anatomy and sexual sensitivity, along with the re‐test reliability of these ratings, in a new self‐reported questionnaire, the Self‐Assessment of Genital Anatomy and Sexual Function, Male (SAGASF‐M).

SUBJECTS AND METHODS

Eighty‐one healthy, sexually active, men aged 22–57 years (median 33), with no history of genital surgery, completed the SAGASF‐M. This questionnaire comprises written text and images enabling men to rate details of their genital appearance, overall genital erotic and pain sensitivity, orgasm intensity, and effort required for achieving orgasm through stimulation of specified areas around the glans and shaft of the penis, scrotum and anus, along with the contribution of other sexually sensitive areas of the body. Anatomical locations were compared for the functional ratings by mixed‐model analysis of variance (anova ). A second sample of 38 healthy men (median age 26 years, range 22–64) from the same source completed the SAGASF‐M twice with an interval of 2 weeks.

RESULTS

There was little variability in anatomy ratings. Ratings of overall penile sensitivity to sexual stimulation gave higher values of ‘sexual pleasure’ for penile stimulation by the partner than by self (P = 0.002) and marginally higher ratings of ‘orgasm intensity’ by partner stimulation (P = 0.077), but there were no corresponding differences on ratings of ‘effort needed to reach orgasm’ or of ‘discomfort/pain’. Overall discrimination between genital areas was highly significant (mixed‐model anova , P = 0.001) for ratings of ‘sexual pleasure’, ‘orgasm intensity’ and ‘orgasm effort’, but was not significant for ‘discomfort/pain’. Ranked by degree of ‘sexual pleasure’, the area ‘underside of the glans’ was highest, followed by ‘underside of the penile shaft’, ‘upper side of the glans’, ‘left and right sides of the glans’, ‘one or both sides of the penis’, ‘upper side of the penile shaft’, ‘foreskin’ (11 subjects), ‘skin between the scrotum and anus’, ‘back side of the scrotum’, ‘front side of the scrotum’, and ‘around anus’, but not all pair differences were significant. The rank order was similar for ‘orgasm intensity’, but less similar and with fewer significant pair differences for ‘orgasm effort’. Overall discrimination of other body parts that help orgasm when touched/stimulated was also highly significant (P = 0.001) and included (in order of degree) scrotum, ear, skin between scrotum and anus, neck, breast/nipples, buttocks, anus (exterior skin), anus (inside with penetration), wrist, and axilla, but many pair differences were not significant. In the reliability study, which was limited to the 45 function items with sufficient variability and sample size, the re‐test reliability values (Pearson r) were distributed as follows: seven were ≥0.80, 16 ≥0.70, 15 ≥0.60, four ≥0.50, two ≥0.40, and one ≥0.30.

CONCLUSION

The SAGASF‐M discriminates reasonably well between various genital and nongenital areas in terms of erotic sensitivity, when administered to genitally unoperated men varying widely in age and socio‐economic level.     

High cytoplasmic expression of p27(Kip1) is associated with a worse cancer-specific survival in clear cell renal cell carcinoma

What's known on the subject? and What does the study add? The loss of p27^Kip1 correlates with poor prognosis in various human cancers, and was postulated as a biomarker in RCC. Up to now p27^Kip1 analysis in RCC was focused on its nuclear localization. We confirmed higher p27^Kip1 expression in the nucleus and cytoplasm of RCC and correlated high cytoplasmic p27^Kip1 with an unfavourable clinic and a reduced survival.

OBJECTIVES

• ? To analyse the cytoplasmic and nuclear differences of p27^Kip1 expression and localization in benign and clear cell renal cell carcinoma (ccRCC) with regard to overall survival (OS) and cancer‐specific survival (CSS).
• ? p27^Kip1 is considered to contribute to the progression of ccRCC and is targeted by next generation dual‐therapies.

PATIENTS AND METHODS

• ? In 140 patients, ccRCC and corresponding benign kidney tissue were analyzed for nuclear and cytoplasmic staining of p27^Kip1 by immunohistochemistry using a tissue microarray technique.
• ? Staining intensity and percentage of positive stained cells are given as expression scores. p27^Kip1 expression was categorized as high if ccRCC tissue stained stronger than the respective level of the corresponding benign tissue and categorized as low if ccRCC tissue stained less than or equal to the corresponding benign tissue.
• ? Differences in OS and CSS were analyzed by log‐rank analysis and expression levels were correlated with tumour and patient characteristics using Fisher's exact test.

RESULTS

• ? Cytoplasmatic (mean [sd ]: 13.8% [1.2%] vs 10.7% [1.7%]; P= 0.04) and nuclear (mean [sd ]: 75.6% [2.7%] vs 13.6% [2.1%]; P < 0.001) staining of p27^Kip1 were significantly stronger in ccRCC tissues compared to benign tissue.
• ? High cytoplasmic p27^Kip1 expression was significantly associated with a higher T and N stage, Fuhrman grade and the presence of metastatic disease (P < 0.001).
• ? The median follow‐up time was 38.2 months.
• ? There was no difference in OS between the low and high expression groups, although CSS was significantly lower in patients with high cytoplasmic p27^Kip1 (P < 0.001) and CSS heavily tended to be lower in the nuclear low expression group (P= 0.069).

CONCLUSIONS

• ? High cytoplasmic p27^Kip1 levels in renal cancer tissues are associated with advanced disease and reduced cancer specific survival, whereas low nuclear expression levels appear to be beneficial.
• ? The present study corroborates the consideration of cytoplasmic p27^Kip1 for future diagnostic and targeted therapeutic approaches in RCC establishing a potential protective shift of p27^Kip1 from the cytoplasm to the nucleus.

     

Prognostic significance of altered p120 ctn expression in bladder cancer

OBJECTIVE

To identify the frequency of change in the expression and localization of p120^ctn in bladder tumours and its association with clinical outcomes, and to investigate the potential role of p120^ctn in the migratory and invasive behaviour of bladder carcinoma cells.

MATERIALS AND METHODS

In all, 425 superficial tumour specimens (Ta, Tis and T1) and 305 invasive (T2–T4) tumour specimens from 534 patients were assembled in 10 tissue microarrays. P120^ctn immunostaining was scored for intensity and cellular localization and correlated with clinical variables and survival analysis. Knockdown of p120^ctn was achieved using small‐interference RNA (siRNA) followed by the assessment of migration and invasion behaviour in standard in vitro assays.

RESULTS

The expression levels of p120 catenin inversely correlated with pathological tumour stage (P < 0.001), histological grade (P < 0.001), presence of lymphovascular invasion (P = 0.02) but not lymph node (LN) involvement (P = 0.17). Non‐membranous localization of p120^ctn correlated with stage (P < 0.001), grade (P < 0.001), lymphovascular invasion (P = 0.04) and LN‐positive disease (P = 0.02). A low expression level of p120^ctn was linked to a poor outcome in cancer‐specific survival analysis. Knockdown of p120^ctn using siRNA resulted in a significant reduction in the migration and invasive potential of bladder carcinoma cells.

CONCLUSIONS

Our findings suggest that p120^ctn acts as a prognostic factor in bladder tumours and has a primary role to play in the migratory and invasive behaviour of bladder carcinoma cells.     

Association of steroid and xenobiotic receptor (SXR) and multidrug resistance 1 (MDR1) gene expression with survival among patients with invasive bladder carcinoma

What’s known on the subject? and What does the study add? SXR and MDR1 are known as responsible for chemo and radiotherapy resistance in some cancers, like kidney cancer (MDR1). Invasive bladder cancer is an aggressive disease, with different behaviour upon its tumoral stage, and also within the same tumoral stage, therefore molecular markers are sought. This study shows a new molecular marker, which has shown as a predictor for bad prognosis cancers, therefore, allowing us for a better patient selection for aggressive therapies.

OBJECTIVE

? To investigate the prognostic value of steroid and xenobiotic receptor (SXR) and multidrug resistance 1 (MDR1) gene expression in relation to survival among patients with invasive bladder cancer.

PATIENTS AND METHODS

? The prospective study included 67 patients diagnosed with invasive bladder cancer and treated with radical cystectomy at one of two institutions. ? SXR and MDR1 gene expression was assessed by real‐time quantitative polymerase chain reaction (RT‐PCR) in tumoral and normal tissue from frozen surgical specimens.

RESULTS

? Patients were followed for a mean of 29 months; 31 patients (46%) had progression. ? In univariate analysis, significant predictors of overall survival (OS) were pathological stage, lymph node (LN) status, histological grade, vascular‐lymphatic invasion, and SXR expression. ? In multivariate analysis, independent predictors of OS were LN status (odds ratio [OR], 2.96; P= 0.034), vascular‐lymphatic invasion (OR, 2.50; P= 0.029), and SXR expression (OR, 1.05, P= 0.03). ? Among the 51 patients with negative LNs (pN0), univariate predictors of OS were SXR expression, MDR1 expression, and pathological stage. In multivariate analysis, SXR expression (OR, 1.06; P= 0.01) and MDR1 expression (OR, 3.27; P= 0.03) were independently associated with survival. ? Within the pN0 group, patients with SXR expression had shorter progression‐free survival than did those without expression (P= 0.004). This association persisted in the N0 subgroup with stage pT3–pT4 disease (P= 0.028). However, in the pN1 group SXR expression did not have any influence.

CONCLUSIONS

? For patients with invasive bladder cancer, SXR expression has value as a predictor of survival independent of the standard pathological predictors. ? Its maximum importance appears to be in patients with stage pT3–pT4 pN0 disease.