Το ουροθηλιακό καρκίνωμα προέρχεται από το ουροθήλιο το οποίο αντιστοιχεί στο επιθήλιο που επενδύει την ουροφόρο οδό. Αποτελεί έναν από τους συχνότερους καρκίνους του ανθρώπου καταλαμβάνοντας την 5η θέση μεταξύ όλων των καρκίνων στο δυτικό κόσμο. Οι σημαντικότεροι παράγοντες κινδύνου για την ανάπτυξη ουροθηλικού καρκινώματος είναι το κάπνισμα και η επαγγελματική έκθεση σε αρωματικές αμίνες. Τα ουροθηλιακά καρκινώματα διακρίνονται σε δύο φαινοτυπικές ποικιλίες με διαφορετική βιολογική συμπεριφορά και πρόγνωση αλλά και διαφορετικούς μηχανισμούς ανάπτυξης. Έτσι έχουμε τα χαμηλού βαθμού κακοήθειας νεοπλάσματα και τους διηθητικούς όγκους (υψηλού βαθμού κακοήθειας). Επιπλέον, ιδιαίτεροι παθογενετικοί μηχανισμοί χαρακτηρίζουν τα ουροθηλιακά νεοπλάσματα της ανώτερης ουροφόρου οδού. Στα χαμηλού βαθμού κακοήθειας νεοπλάσματα τα οποία προέρχονται από υπερπλαστικές αλλοιώσεις συναντάμε μεταλλάξεις των υποδοχέων FGFR3 και EGFR καθώς και του ογκογονιδίου H-RAS με αποτέλεσμα την ενεργοποίηση του μονοπατιού των RAS-MAP κινασών. Στους υψηλού βαθμού κακοήθειας (διηθητικους) όγκους οι οποίοι αναπτύσσονται de novo ή προέρχονται από καρκίνωμα in situ συναντάμε μεταλλάξεις των ογκογονιδίων p53, RB και p57. Επιπλέον χαρακτηρίζονται από υπερέκφραση της αντιαποπτωτικής πρωτεϊνης survivin, των καβεολινών 1 και 2, της COX-2, των μεταλλοπρωτεϊνασών και των επαγωγέων της αγγειογένεσης VEGF, VEGFR και bFGF, καθώς και από διαταραχή της έκφρασης των μορίων του κυτταροσκελετού. Τέλος έχει βρεθεί υπερέκφραση του παράγοντα OCT-4, ρυθμιστή της αυτοανανέωσης και της διαφοροποίησης στα εμβρυϊκά stem κύτταρα, δικαιώνοντας τη θεωρία των καρκινικών stem κυττάρων. Η απώλεια ετεροζυγωτίας του χρωμοσώματος 9 παίζει επίσης σημαντικό ρόλο στη ανάπτυξη ουροθηλιακών καρκινωμάτων και θεωρείται ότι εμπλέκεται στα πρώιμα στάδια της ογκογένεσης απαντώμενη έτσι και στις δύο φαινοτυπικές ποικιλίες. Αντίθετα, τα ουροθηλιακά νεοπλάσματα της ανώτερης ουροφόρου οδού χαρακτηρίζονται από ένα είδος γενετικής αστάθειας, την αστάθεια των μικροδορυφόρων. Η καλύτερη κατανόηση των παθογενετικών μηχανισμών ανάπτυξης των ουροθηλιακών καρκινωμάτων θα συμβάλλει τόσο στη δημιουργία νέων και αποτελεσματικότερων φαρμάκων όσο και στην εύρεση νέων μοριακών δεικτών. Ο προσδιορισμός τέτοιων βιολογικών δεικτών, θα βελτιώσει το screening και τη διάγνωση και θα βοηθήσει στον καλύτερο προσδιορισμό του κακοήθους δυναμικού αλλά και της πρόγνωσης των ουροθηλιακών καρκινωμάτων. / Urothelial cancer derives from urothelium which is the epithelium lining the urinary tract. It is one of the most frequent human cancers occupying the 5th position among all cancers in the Western World. The most important risk factors for the development of urothelial cancer are smoking and occupational exposure to aromatic amines. Urothelial cancers are divided into two dinstict categories according to their biological behaviour and pathogenetic background: low grade neoplasms and high grade tumors (infiltrative). Interestingly, the neoplasms of the upper urinary tract have a distinct pathogenetic mechanism. Low grade urothelial carcinomas, known to derive from hyperplastic lesions, have been found to bear mutations of the FGFR3 and EGFR receptors as well as of the H-RAS oncogene which leads to the activation of the RAS-MAPK signal transduction pathway. High grade tumors may develop de novo or they may derive from carcinoma in situ. They are commonly characterized by mutations of the oncogenes p53, Rb and p57. Additionaly, in high grade urothelial carcinomas there is overexpression of the antiapoptotic protein survivin as well as caveolin1 and 2, COX-2, metalloproteases and angiogenetic factors such as VEGF, VEGFR and bFGF. There is also disrupted expression of the cytoskeleton’s molecules. Overexpression of the transcription factor OCT-4, a regulator of self renewal and differentiation of embryonic stem cells, has been also recently reported. Loss of heterozygosity of chromosome 9 plays an important role in the development of urothelial cancer and it is considered to be implicated in the early steps of oncogenesis. In contrast, urothelial cancers of the upper urinary tract have been associated with microsatellite instability, an indicative marker of genomic instability. Elucidating the pathogenesis of urothelial cancersmay contribute to the development of novel and more effective anticancer treatments as well as to the identification of new biological markers with prognostic significance.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23565号 / 医博第4779号 / 新制||医||1054(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 武藤 学, 教授 万代 昌紀, 教授 上杉 志成 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Les carcinomes urothéliaux représentent la 4ème cause de cancer chez l'homme après les cancers de la prostate, du colon et du poumon. Leur incidence est en augmentation de plus de 50% depuis 25 ans. Ce cancer présente principalement deux formes, une superficielle (70% des cas) de bon pronostic et une invasive (30%) de mauvais pronostic. Les formes superficielles nécessitent une surveillance active rapprochée afin d'identifier les récidives fréquentes et l'évolution vers un stade invasif. Cette surveillance fait principalement appel à la cystoscopie et engendre morbidité et cout importants. Une alternative à la cystoscopie est possible à l'aide de tests de détection urinaire des cellules cancéreuses, mais leur sensibilité jusqu'à présent n'est pas suffisante pour une utilisation en pratique courante. Notre but a été de développer un outil de détection urinaire des tumeurs urothéliales.Matériel, Méthode et RésultatsLes tumeurs urothéliales étant très instables sur le plan génétique, le travail a initialement consisté à faire la synthèse des anomalies cytogénétiques retrouvées dans la littérature. Les anomalies les plus pertinentes ont été sélectionnées et une puce de CGH en a été développée comprenant 341 clones (BAC) répartis sur l'ensemble des chromosomes. Cette puce intitulée BCA-1 a été développée en collaboration avec la société ArrayGenomics (Voisins Le Bretonneux, France). La validité de ce test a été confirmée sur 10 lignées cellulaires tumorales et bénignes.Notre travail a ensuite consisté à étudier la valeur ajoutée de cette puce en pratique clinique. Pour ce faire, une cohorte de 163 patients porteurs de tumeurs urothéliales et de témoins a été constituée. Les urines ont été prélevées et analysées en utilisant la puce BCA-1. Un logiciel a été développé sous filemaker pro afin de permettre une saisie uniforme et détaillée des données cliniques et de prendre en considération le caractère complexe de la prise en charge de ces tumeurs.Le test urinaire utilisant la puce de CGH a montré une excellente performance diagnostique avec une sensibilité de 96% et une spécificité de 98% dans les tumeurs vésicale, et une sensibilité de 100% tumeurs du haut appareil urinaire.Enfin, le test a aussi permit de caractériser le caractère agressif ou non agressif des tumeurs sur le plan cytogénétique. Cette caractérisation est fortement corrélée avec le stade anatomopathologique et un troisième aspect de notre travail a montré que la détermination cytogénétique de l'agressivité des tumeurs prédisait l'évolution défavorable des tumeurs du haut appareil urinaire. Notre travail a permis le développement et l'analyse d'un nouveau test de dépistage urinaire des tumeurs urothéliales permettant le diagnostic urinaire de ces tumeurs et la caractérisation de leur caractère agressif éventuel, ainsi que le développement d'un logiciel de saisie et d'analyse des données cliniques. / Urothelial carcinomas are the 4th cause of cancer in men, following prostate, colon and lung cancer. An increase of 50% in its incidence was observed on the last 25 years. This cancer presents two types, a superficial type (70% of the cases) of good prognosis and an invasive type (30% of the cases) of bad prognosis. Superficial type require an active monitoring to identify recurrences and evolution to an invasive stage. This follow up is performed using cystoscopy and leads to some level of morbidity and a high cost. An alternative to cystoscopy is possible using urine test to detect cancer cells, but they are lacking of sensitivity to be used instead of cystoscopy in clinical practice. Our goal was to develop a urine detection tool of urothélial carcinomas.Material and MethodsUrothelial carcinoma usually present with a high level of genetic instability. We first analyse literature so to identify most relevant cytogenetic abnormalities that occur in urothélial carcinomas. A CGH array chip was designed using 341 clones (BAC) that were selected according to initial analysis. This chip called BCA-1 covers the whole genome and was developed in collaboration with ArrayGenomics (Voisins Le Bretonneux, France).ResultsThis test has shown a good efficacy on a preliminary study of cytogenetic analysis on 10 cancerous and benign bladder cell lines. The Chip was then assessed in clinical practice on a series of 163 patients diagnosed with or without urothelial cancer. Urines were collected and analysed using the BCA-1 chip. A software was designed to favour homogeneous and detailed clinical data collection and take into consideration the complex management of the tumours.The test using the CGH chip as shown an excellent diagnosis performance with a sensitivity of 96% and a specificity of 98% for bladder cancer detection, and a sensitivity of 100% on upper urinary tract detection.Finaly, the test was able to define the grade of the tumour according to cytogenetic loci affected. This grade was strongly correlated with the pathology score and could be used to predict outcomes in upper urinary tract carcinomas. Our work lead to the developpement and the analysis of a new urothélial carcinoma urinary detection test, to the identification of the agressivity of the tumour, and to the development of an analysis and data entry software for clinical details.

Epidemiologic data suggest that soy consumption may protect against cancer induction in several tissues in humans, including urothelial carcinoma. Genistein have been reported to regulate genes that are involved in several cellular events. However, the molecular mechanism of genistein -induced upregulation of epithelial membrane protein 2 (EMP2), candidate urothelial tumor suppressor, is not entirely understood. At first, we found that the mRNA and protein expression levels of EMP2 were significantly greater in the normal urothelial tissues and human urothelial cells than those in urothelial bladder carcinoma tissues and urothelial cell carcinoma-derived cell lines. Second, EMP2 knockdown via RNA interference markedly enhanced cell proliferation, colony formation, migration and invasiveness. By contrast, EMP2 overexpression suppressed these malignant behaviors. Third, we showed that genistein-induced inhibition in cell proliferation is associated with an increase in EMP2 expression. Using various deleted EMP2 promoter constructs, we defined that the EMP2 core promoter is enough to observe the genistein-induced upregulation of EMP2 transcriptional activity. Using site direct mutagenesis and chromatin immunoprecipitation assays demonstrated that cyclic-AMP response element binding protein 1 (CREB1) acts as a positive regulator of EMP2 transcription by directly binding to its promoter. These results showed EMP2 suppressed urothelial cell carcinoma-derived cell growth, motility and invasion and for the first time that genistein promoted EMP2 expression in urothelial cell carcinoma-derived cells by inducing EMP2 transcriptional activity via CREB1 binding.

Purpose: To determine the expressions of bcl-2, p53, Ki-67 and PTEN on the basis of immunohistochemistry methods in upper urinary transitional cell carcinoma (TCC), and to correlate their presentations in specimens with clinical tumor stage, grade and patient survival. Material and Method: Paraffin-embedded primary upper urinary TCC specimens were divided into 2 groups for immunohistochemical study: Group 1 including 91 cases were treated with bcl-2, p53 and Ki-67 antibodies; group 2 including 93 specimens contained both tumor and benign tissues were treated with PTEN antibody. Semi- quantitatively, according to the amount of the stained cells, they were divided into 3 levels: level 1, scanty; level 2, focal; and level 3, diffuse. Association of immunoreactivity with tumor grade and stage was examined. Prognostic significance of tumor marker expression in patients¡¦ survival was accessed. Results: Group1: Of the 91 tumors most (98.9%) of the specimens showed level 1 bcl-2 expression and only 1 patient had level 2 expression. The p53 mutations were identified level 3 expression in 48.4% of the cases, followed by level 2 (26.4%) and level 1 (25.3%) identifications. The Ki-67 expression was recognized level 3 in 6 patients, level 2 in 21 and level 1 in 66 cases. Significant correlations were seen between p53 expression and tumor grading (p=0.004) and between immunostain of Ki-67 and clinical stage (p=0.031). The p53, bcl-2 and Ki-67 expressions in upper urinary tract TCC specimens were not a significant factor of patients¡¦ survival. Group 2: Of the tumors all cytoplasm has level 3 PTEN expressions and the nuclei, 18 (19.4%) showed scanty expression, 35 (37.6%) revealed focal expression, and diffuse expression was noted in 40 (43.0%) cases. Loss of PTEN expression in tumor nuclei was positively correlated with pathologic stage (p=0.019). Of the fibrocytes adjacent to tumor cells, the nuclei showed 24 (25.8%) scanty, 59 (63.4%) focal and 10 (10.8%) diffuse distribution of PTEN expressions. Poorly differentiated tumor (grade 3) specimens were correlated with loss of PTEN expression in fibrocytic nuclei adjacent to tumor (p=0.028). Most (58%) fibrocytic cytoplasm was scanty PTEN expression, followed by 23 (24.7%) diffuse and 16 (17.2%) focal immunostaining. PTEN expressions in upper urinary tract TCC specimens were not a significant factor of patients¡¦ survival. Conclusions: We examined 93 surgical specimens of upper urinary tract TCC for the expression of PTEN and 91 cases for bcl-2, p53 and Ki-67 by immunohistochemical stained. Correlation between tumor grading and p53 mutations and correlation between clinical stage and Ki-67 immunoreactivity were observed. Meanwhile, loss of PTEN expression in tumor nuclei of upper urinary TCC is correlated significantly with advanced tumor stage, and poorly differentiated tumor specimens were correlated with loss of PTEN expression in normal nuclei adjacent to tumor cells. However, no correlation between overall survival rate and tumor markers was identified. Thus, the detection of p53, bcl-2, Ki-67 and PTEN would be not enough for evaluation the prognosis of upper TCC.

Bladder cancer has been associated with chronic arsenic exposure. Monomethylarsonous acid [MMA(III)] is a metabolite of inorganic arsenic biotransformation and has been shown to transform an immortalized urothelial cell line (UROtsa) at a concentration 20-fold less than arsenite. MMA(III) was used as a model arsenical to examine the mechanisms of arsenical-induced transformation of the urothelium. A microarray analysis was performed to assess the transcriptional changes in UROtsa during the critical window of chronic MMA(III) exposure that leads to transformation at three months time. The analysis revealed only minor changes in gene expression at one and two months of exposure, contrasting with substantial changes observed at three months of exposure. The gene expression changes at three months were analyzed showing distinct alterations in biological processes and pathways such as a response to oxidative stress, enhanced cell proliferation, anti-apoptosis, MAPK signaling, as well as inflammation. To address the lack of information between two and three months of exposure -- the critical period of transformation -- the expression of selected pathway marker genes were measured by PCR array analysis on a weekly and monthly basis. A very similar pattern of altered expression of these genes was observed when compared to microarray results, and suggested early perturbations in cell signaling cascades, immunological pathways, cytokine expression, and MAPK pathway, are particularly important in driving malignant transformation. These results showed a strong association between the acquired phenotypic changes that occurred as early as one to two months of chronic MMA(III) exposure, and gene expression patterns that are indicative of the earliest stages in carcinogenesis. Additionally, studies on the effects of withdrawal of arsenical were also conducted and showed that phenotypic changes persisted even in the absence of arsenical; that gene expression patterns of pathway marker genes, those that showed significant alterations between 3 and 6 months of exposure, appeared to normalize after withdrawal of the arsenical.

Background: Lynch syndrome is a hereditary disorder, with a very high risk of the developing colorectal cancer (CRC) and a predilection to develop other cancers, including upper tract urothelial carcinoma (UTUC) that has an estimated lifetime risk of 0.2-25%, above that of the general population. Our aim was to assess the prevalence of UTUC in a Lynch syndrome cohort undergoing screening for CRC, to determine the need for a UTUC screening program. Methodology: Lynch syndrome patients were screened with urine dipstix for microscopic haematuria. Patients with confirmed microhaematuria were offered urine cytology, microscopy and culture, ultrasound (US) of their upper tracts and flexible cystoscopy. Results: Of the 89 patients screened, 86 had an MLH1 mutation and 2 had an MSH2 mutation. Eleven of the 12 patients who had microscopic haematuria were female. 10 patients had urinary tract infections. One patient had follicular cystitis and another had a simple renal cyst. No patients had hydronephrosis on ultrasound. All urine cytology specimens were negative for malignancy. Conclusion: No cases of UTUC were detected in our cohort during this study. A more rational screening protocol in this group may be to screen patients for UTUC with known MSH2 mutations at an earlier age (over 35).

Affibodymolekyler är små affinitetsproteiner (6.5 kDa) som föreslås kunna ersätta monoklonala antikroppar i terapeutiska tillämpningsområden, exempelvis i antikropp-läkemedelskonjugat (ADCs) som kan navigera sig fram till biomarkörer som är uttryckta på cancerceller. Affibody-läkemedelskonjugat (AffiDC) kan användas för att målsöka just sådana överuttryckta proteiner, samtidigt som de erbjuder goda egenskaper, såsom snabb transportering och spridning i kroppen, och effektiv penetrering genom tumörer. Dessa AffiDC:er skulle kunna användas inom riktad cancerterapi for de cancersjukdomar som fortfarande är i behov av cancerhämmande behandlingar, såsom urotelial cancer.  Den här studien föreslog tillämpning av ABD-kopplade affibodymolekyler för att målsöka ett nytt målprotein som har visats vara överuttryckt i flera olika cancersjukdomar, exempelvis bröst-, pankreas- och urotelial cancer. Affibodykandidater mot målproteinet har valts ur ett rekombinant bibliotek med 1×1011 transformanter som uttrycks med hjälp av en så kallad metod med E. coli celldisplay där affibodymolekylen visas på cellens ytmembran. De slutliga kandidaterna var sedan identifierade och biokemiskt karaktäriserade i in vitro-studie på människoceller, som visade att två av kandidaterna verkade binda till cancercellinjerna BT-474 och MCF-7 med KD omkring 10 till 100 nM. / Affibody molecules are small affinity proteins (6.5 kDa) suggested to substitute monoclonal antibodies in therapeutic applications, e.g., antibody-drug conjugates (ADCs) targeting biomarker proteins expressed on cancer cells. An affibody-drug conjugate (AffiDC) could be used to target these types of overexpressed proteins on cancer cells while offering attractive properties, such as rapid transportation and distribution in the body, as well as efficient tumour penetration. These AffiDCs could be used as a targeted cancer therapy for cancers that are yet to be treatable and curable, like urothelial cancers.  This study suggested the use of ABD-fused affibodies to target a novel cancer protein that has been shown to be overexpressed on cancer cells, including breast, pancreatic and urothelial cancer. Affibody candidates toward this novel target were selected from a recombinant library, of 1×1011 transformants, that is expressed using E. coli cell display system. The final candidates were subsequently biochemically characterized and assessed for affinity for the target. Three affibodies were finally identified and assessed in in vitro studies on mammalian cells, revealing two affibodies that appear to bind to the cell lines BT-474 and MCF-7 with KD ranging 10 to 100 nM.

Objetivo: Identificar os principais fatores prognósticos patológicos do carcinoma urotelial tratado com cistectomia radical e linfadenectomia pélvica, analisar o impacto desses na sobrevida livre de doença (RFS), câncer específico (CSS), geral (OS) e propor um conjunto de fatores que possa prever a evolução. Métodos: Realizamos levantamento dos casos de cistectomia radical e linfadenectomia no período de 2006 a 2009 no Hospital das Clinicas da FMUSP. Correlacionamos fatores prognósticos patológicos estádio (pT), grau tumoral, presença de metástase linfática (pN), invasão linfovascular (LVI), perineural (PNI) e presença de CIS com RFS, CSS e OS. Consideramos significante quando p > 0,05. Resultados: Avaliamos 128 casos que obedeceram aos critérios sendo 20 (15,6%) femininos e 108 (84,4%) masculinos, idades variando de 41 a 84 anos e média de 67 anos. Constatamos associação de recidiva com estádio >pT2 (p=0,032) e pN+ (p=0,003). Os fatores estádio >pT2 (p=0,001), pN+ (p=0,034) LVI+ (p=0,038), e PNI+ (p=0,024) tiveram associação com óbito pela doença e a morte geral com estádio >pT2 (p=0,001), pN+ (p=0,038) e PNI+ (p=0,01). A análise multivariada demonstrou que apenas estádio >pT2 e pN+ são os fatores prognósticos independentes na RFS, CSS e OS. Conclusão: A análise dos fatores patológicos após cistectomia radical e linfadenectomia demonstrou que estádio >pT2 e pN+ são os fatores prognósticos mais importantes no CaB / Objective: To identify the most important pathologic prognostic factors for urothelial bladder cancer treated by cystectomy and pelvic lymphadenectomy, analyze the impact of these on recurrence and mortality and suggest a group of factors that can predict the outcome after surgery. Method: We review all radical cystectomy and lymphadenectomy cases at the Clinical Hospital of São Paulo Medical School from 2006 to 2009. We correlate the following pathologic prognostic factors tumor stage (pT), tumor grade, lymphonodal metastasis (pN), lymphovascular invasion (LVI), perineural invasion (PNI), presence of CIS with RFS, CSS and OS. We considered a significant association p<0.05. Results: We selected 128 cases for this study. There were 20 (15.6%) females and 108 (84.4%) males, ages ranging from 41 years to 84 years and an average 67 years old. The tumor recurrence was associated with stage >pT2 (0,032) and pN+ (p=0.003). Stage >pT2 (p=0.001), pN+ (p=0.034), LVI+ (p=0.038) and PNI+ (p=0.024) was associated with death by cancer and overall death with stage >pT2 (p=0.001), N+ (p=0.038) and PNI (p=0.01). The multivariate analysis found that only stage >pT2 and pN+ were independent prognostic variable for RFS, CSS and OS. Conclusion: The analysis of pathologic prognostic factors after radical cystectomy and pelvic lymphadenectomy show that stage >pT2 and pN+ has strong association with RFS, CSS and OS

Introdução: O carcinoma urotelial de bexiga (CUB) é o segundo tumor mais frequente do trato urinário. A perda da função do p53 é a alteração mais conhecida do carcinoma urotelial de alto grau invasivo. Prima-1 é uma pequena molécula, a qual restaura a função do p53 mutado promovendo a morte celular em vários tipos celulares. O objetivo do nosso estudo foi analisar o efeito do Prima-1 na indução da apoptose mediada por p53 após indução do dano ao DNA em linhagens de CUB. Material e métodos: O mecanismo de ação do Prima-1 foi avaliado em duas linhagens celulares de câncer de bexiga, T24 que tem como característica a mutação do p53 e RT4 que possui p53 intacto. Características morfológicas da apoptose, alterações no potencial de membrana mitocondrial e análise da expressão de treze genes envolvidos na apoptose mediada por p53 foram avaliados através de observação microscópica e reação em cadeia da polimerase em tempo real quantitativa (qRT-PCR). Resultados: Prima-1 foi capaz de reativar a função da P53 na linhagem de câncer de bexiga com p53 mt, promovendo a apoptose através da expressão de Bax e Puma, ativação da cascata das caspases e ruptura da membrana mitocondrial, independente de Bax, na linhagem celular T24 (p53 mt). Conclusão: Prima-1 foi capaz de restaurar a atividade transcricional de p53. Estudos experimentais in vivo poderiam ser conduzidos no intuito de testar essa molécula como um novo agente terapêutico do CUB de alto grau, invasivo, que apresenta caracteristicamente mutação de p53 / Introduction: Urothelial carcinoma of the bladder is the second most common tumor of the urinary tract. Loss of p53 function is the main genetic alteration related to the development of high-grade muscle-invasive disease. Prima-1 is a small molecule that restores tumor suppressor function to mutant p53 and induces cell death in various cancer types. Our aim is to investigate the ability of Prima-1 to induce apoptosis after DNA damage in BC cancer cell lines. Material and Methods: The therapeutic effect of Prima-1 was studied in two BC cell lines, T24, characterized by p53 mutation, and RT4, with no mutation in the p53 gene. Morphological features of apoptosis, mitochondrial membrane potential changes and expression of thirteen genes involved in p53-induced apoptosis were assessed by microscopic observation and quantitative real-time PCR (qRT-PCR) Results: Prima-1 is able to reactivate P53 function in p53-mutated bladder cancer cell line promote apoptosis through the induction of Bax and Puma expression, activating the caspase cascade and disruption of mitochondrial membrane, independent of Bak, in T24 cell line (p53 mt). Conclusion: Prima-1 is able to restore the transcriptional activity of p53. Experimental studies in vivo could be conducted in order to test this molecule as a new therapeutic agent of the urothelial carcinoma of the bladder, which characteristically presents p53 mutation