Stéphane Larré

Distinct MicroRNA Alterations Characterize High- and Low-Grade Bladder Cancer

Urothelial carcinoma of the bladder (UCC) is a common disease that arises by at least two different molecular pathways. The biology of UCC is incompletely understood, making the management of this disease difficult. Recent evidence implicates a regulatory role for microRNA in cancer. We hypothesized that altered microRNA expression contributes to UCC carcinogenesis. To test this hypothesis, we examined the expression of 322 microRNAs and their processing machinery in 78 normal and malignant urothelial samples using real-time rtPCR. Genes targeted by differentially expressed microRNA were investigated using real-time quantification and microRNA knockdown. We also examined the role of aberrant DNA hypermethylation in microRNA downregulation. We found that altered microRNA expression is common in UCC and occurs early in tumorogenesis. In normal urothelium from patients with UCC, 11% of microRNAs had altered expression when compared with disease-free controls. This was associated with upregulation of Dicer, Drosha, and Exportin 5. In UCC, microRNA alterations occur in a tumor phenotype-specific manner and can predict disease progression. High-grade UCC were characterized by microRNA upregulation, including microRNA-21 that suppresses p53 function. In low-grade UCC, there was downregulation of many microRNA molecules. In particular, loss of microRNAs-99a/100 leads to upregulation of FGFR3 before its mutation. Promoter hypermethylation is partly responsible for microRNA downregulation. In conclusion, distinct microRNA alterations characterize UCC and target genes in a pathway-specific manner. These data reveal new insights into the disease biology and have implications regarding tumor diagnosis, prognosis and therapy.

Combination of Polymorphisms From Genes Related to Estrogen Metabolism and Risk of Prostate Cancers: The Hidden Face of Estrogens

PURPOSE The association between common functional polymorphisms from the CYP17, CYP19, CYP1B1, and COMT genes involved in the estrogen metabolism and the risk of prostate carcinoma was evaluated. PATIENTS AND METHODS The study investigated 1,983 white French men (1,101 patients with prostate cancer and 882 healthy controls) aged between 40 and 98 years. The different alleles and genotypes were analyzed according to case-control status, aggressiveness pattern of the tumors, age at onset, and family history of cancers. RESULTS The VV (high activity) genotype of the V432L polymorphism from CYP1B1 (odds ratio [OR] = 1.36; 95% CI, 1.03 to 1.79; P = .031), and the long allele (> 175 bp) of the TTTA repeat from CYP19 (OR, 1.26; 95% CI, 1.08 to 1.47; P = .003) were significantly associated with the risk of prostate cancer. An additive effect was observed when we combined the two at-risk alleles (OR = 1.63; 95% CI, 1.24 to 2.13; P < .001). The association was stronger for the CYP1B1 VV genotype (OR = 1.55; 95% CI, 1.13 to 2.13; P = .007) among the group of patients with highly aggressive disease. Stratification by age at onset showed that the associations of CYP1B1 and CYP19 variants were largely confined to the younger prostate cancer patients. CONCLUSION This association between polymorphisms from genes related to estrogen metabolism and prostate cancer risk suggest new clinical considerations in the management of prostate cancer: the development of new prevention trials based on genetic profiling and the evaluation of specific inhibitors involving the estrogen pathways.

Recommandations en Onco-Urologie 2010 : Tumeurs urothéliales

.Tx Tumeur primitive non evaluable .T0 Tumeur primitive non retrouvee .Ta Carcinome papillaire non invasif .Tis Carcinome in situ « plan » .T1 Tumeur envahissant le chorion .T2 Tumeur envahissant la musculeuse – T2a Tumeur envahissant le muscle superficiel (moitie interne) – T2b Tumeur envahissant le muscle profond (moitie externe) .T3 Tumeur envahissant le tissu peri-vesical – T3a Envahissement microscopique – T3b Envahissement extra-vesical macroscopique .T4 Tumeur envahissant une structure peri-vesicale – T4a Prostate, vagin ou uterus – T4b Paroi pelvienne ou abdominale

S100A6 (Calcyclin) is a prostate basal cell marker absent in prostate cancer and its precursors

S100A6 (Calcyclin) is a calcium-binding protein that has been implicated in a variety of biological functions as well as tumorigenesis. The aim of our study was to investigate the involvement of S100A6 during prostate cancer development and progression. Using immunohistochemistry, the expression of S100A6 was examined in benign (n=66), premalignant (n=10), malignant (n=66) and metastatic prostate (n=5) tissues arranged in a tissue-microarray or whole sections as well as in prostate cancer cell lines. The S100A6 immunostaining pattern in tissues was compared with that of cytokeratin 5 (a basal cell marker) and 18 (a benign luminal cell marker). In all cases of benign epithelium, intense S100A6 expression was seen in the basal cell layer with absent staining in luminal cells. In all cases of prostatic adenocarcinoma (matched), metastatic lesions and 3/10 high-grade prostatic intraepithelial neoplasia lesions, an absence of S100A6 was seen. Western blotting and RT–PCR analysis of cell lines showed S100A6 expression to be absent in LNCaP, LNCaP-LN3 and LNCaP-Pro5 but present in Du145, PC3, PC-3M and PC-3M-LN4. LNCaP cells treated with 5-Azacytidine, caused re-expression of S100A6 mRNA. Sequencing of bisulphite modified DNA showed CpG methylation within the S100A6 promoter region and exon 1 of LNCaP, LNCaP-LN3 and LNCaP-Pro5 cell lines but not in Du145 cells. Our data suggest that loss of S100A6 protein expression is common in prostate cancer development and may occur at an early stage. The mechanism of loss of expression may involve hypermethylation of CpG sites. The finding of intense S100A6 expression in the basal cells of benign glands but loss of expression in cancer could be useful as a novel diagnostic marker for prostate cancer.

Screening for Bladder Cancer: Rationale, Limitations, Whom to Target, and Perspectives

CONTEXT Bladder cancer (BCa) is the fourth most common cancer in men. Survival from the disease has not improved in the last 25 yr. Population-based screening theoretically provides the best opportunity to improve the outcomes of aggressive BCa. OBJECTIVE To review the current literature regarding the usefulness and feasibility of screening for bladder cancer. EVIDENCE ACQUISITION We conducted a nonsystematic review restricted to English using the keywords urinary bladder neoplasms, mass screening, mandatory testing, and early detection of cancer. We retrieved 184 articles and selected 22. EVIDENCE SYNTHESIS There was no level 1 evidence (obtained from a randomised controlled trial [RCT]) addressing the impact of screening on BCa survival or tumour downstaging. No study assessed the diagnostic performance of urinary markers in the context of screening. Two case-control series suggested a benefit of screening on survival, and a third found a nonsignificant beneficial trend in favour of screening. Two studies suggested downstaging of BCa at diagnosis. Other reports concluded that most cancers detected with screening were of low grade and that current urinary testing cannot detect all tumours. Screening is likely to be of benefit in high-risk populations using cost-efficient high-performing urinary biomarkers. There was insufficient evidence to define an efficient screening protocol. CONCLUSIONS Although BCa screening is theoretically feasible in a high-risk population, there is currently insufficient evidence to recommend it. This is due to insufficient data to define an efficient screening protocol with selection of an appropriate population and the lack of accurate and cost-effective urinary markers able to discriminate low-risk from high-risk cancers. Major improvements are needed in the evaluation of urinary biomarkers before evaluation in a RCT can be achieved.

PGE2 and LTB4 tissue levels in benign and cancerous prostates.

PGE2 and LTB4 are involved in inflammation and carcinogenesis in several tissues but have not been studied in prostate cancer and hyperplasia until now. We therefore measured PGE2 and LTB4 productions in a total of 206 prostate tissues from 116 patients including benign hyperplastic (90), pericancerous (106) and cancerous samples (10). We also analysed the influence of inflammation levels, prostate volume and glandular to epithelial ratio. PGE2 and LTB4 concentrations were measured using specific enzyme immunoassay kits. There was a correlation between PGE2 level, prostatic volume, inflammation score, and decreased glandular surface. By contrast, there was no correlation between LTB4 levels and inflammation or PGE2 production. Cancerous samples had higher LTB4 levels than pericancerous samples, but there was no difference in PGE2 levels. PGE2 and inflammation may be associated to stromal benign prostatic hyperplasia whereas LTB4 may play a role in prostate carcinogenesis.

Prognostic Interest in Discriminating Muscularis Mucosa Invasion (T1a vs T1b) in Nonmuscle Invasive Bladder Carcinoma: French National Multicenter Study with Central Pathology Review

PURPOSE Predictive factors of T1 nonmuscle invasive bladder cancer evolution that could guide treatment decision making are lacking. We assessed the prognostic value of muscularis mucosa invasion in nonmuscle invasive bladder cancer. MATERIALS AND METHODS In a national multicenter study patients with primary T1 nonmuscle invasive bladder cancer were recruited from 6 French hospitals. All patients had undergone transurethral resection of bladder tumor. All T1 tumors were substaged according to muscularis mucosa invasion as T1a-no invasion beyond the muscularis mucosa or T1b-invasion beyond the muscularis mucosa with muscle preservation. Subsequent central pathology review was then done by a single referent uropathologist. Muscularis mucosa invasion was tested as a prognostic factor for survival on univariate and multivariate analysis. RESULTS A total of 587 patients were enrolled in the study, including 388 (66%) with T1a and 199 (34%) with T1b tumors. Median followup after transurethral resection of bladder tumor was 35 months (IQR 14-54). There was no significant difference between groups T1a and T1b except high tumor grade in T1b cases (p <0.0001). After central review, initial pathological substaging was confirmed in 84% of cases. On multivariate analysis muscularis mucosa invasion (T1b substage) was significantly associated with recurrence-free (p = 0.03), progression-free (p = 0.0002) and cancer specific (p = 0.02) survival. The main study limitation was absent systematic subsequent transurethral resection of bladder tumor. CONCLUSIONS Muscularis mucosa invasion appears to be highly predictive of T1 nonmuscle invasive bladder cancer behavior. Consequently, systematic T1a vs T1b discrimination should be highly advocated by urologists and pathologists. We believe that it could aid in crucial decision making when choosing between conservative management and radical cystectomy remains a moot point.

Phenol Sulfotransferase SULT1A1*2 Allele and Enhanced Risk of Upper Urinary Tract Urothelial Cell Carcinoma

Cytosolic sulfotransferases (SULT) are involved in detoxification pathways. A functional polymorphism in the SULT1A1 gene, leading to an Arg213His substitution (SULT1A1*2), is thought to confer susceptibility to various types of cancer. Upper urinary tract urothelial cell carcinomas (UUT-UCC) are rare (5% of all urothelial carcinomas). We genotyped 268 patients with UUT-UCC and 268 healthy controls matched for age, gender, tobacco consumption, and ethnicity. His213 (SULT1A1*2) allele frequency was significantly higher in patients than in controls (37.1% versus 28.9%; P = 0.004). The His/His genotype corresponding to low-activity SULT1A1 enzyme conferred a significantly higher risk of UUT-UCC (odds ratio, 2.18; 95% confidence interval, 1.28-3.69; P = 0.004).(Cancer Epidemiol Biomarkers Prev 2007;16(11):1–4) (Cancer Epidemiol Biomarkers Prev 2007;16(11):2500–3)

Neurofuzzy Modeling to Determine Recurrence Risk Following Radical Cystectomy for Nonmetastatic Urothelial Carcinoma of the Bladder

Purpose: Bladder cancer recurrence occurs in 40% of patients following radical cystectomy (RC) and pelvic lymphadenectomy (PLND). Although recurrence can be reduced with adjuvant chemotherapy, the toxicity and low response rates of this treatment restrict its use to patients at highest risk. We developed a neurofuzzy model (NFM) to predict disease recurrence following RC and PLND in patients who are not usually administered adjuvant chemotherapy. Experimental Design: The study comprised 1,034 patients treated with RC and PLND for bladder urothelial carcinoma. Four hundred twenty-five patients were excluded due to lymph node metastases and/or administration of chemotherapy. For the remaining 609 patients, we obtained complete clinicopathologic data relating to their tumor. We trained, tested, and validated two NFMs that predicted risk (Classifier) and timing (Predictor) of post-RC recurrence. We measured the accuracy of our model at various postoperative time points. Results: Cancer recurrence occurred in 172 (28%) patients. With a median follow-up of 72.7 months, our Classifier NFM identified recurrence with an accuracy of 0.84 (concordance index 0.92, sensitivity 0.81, and specificity 0.85) and an excellent calibration. This was better than two predictive nomograms (0.72 and 0.74 accuracies). The Predictor NFMs identified the timing of tumor recurrence with a median error of 8.15 months. Conclusions: We have developed an accurate and well-calibrated model to identify disease recurrence following RC and PLND in patients with nonmetastatic bladder urothelial carcinoma. It seems superior to other available predictive methods and could be used to identify patients who would potentially benefit from adjuvant chemotherapy.

The outcome of patients with pathological Gleason score ≥8 prostate cancer after radical prostatectomy

To analyse the outcome of patients undergoing radical prostatectomy (RP) for Gleason 8–10 clinically localized prostate cancer, and to evaluate the prognostic value of well‐known predictors of progression.