Boosting bladder cancer treatment by intravesical nitazoxanide and bacillus calmette-guérin association.

PURPOSE: Nitazoxanide (NTZ) has shown a promising antitumoral effect, the current study compared the anti-neoplastic effects of intravesical NTZ and BCG plus NTZ in NMIBC animal model. METHODS: 30 rats, Fisher 344 were instilled with 4 intravesical doses of 1.5 mg/kg of N-methyl-N-nitrosourea (MNU) every 15 days for BC induction. The animals were divided into 3 groups (Group BCG 106 UFC - 1 mg of BCG; Group NTZ - 300 mg/kg of NTZ; Group NTZ + BCG - simultaneous treatment of BCG and NTZ) and received weekly intravesical treatment for 6 consecutive weeks. Animals were submitted to ultrasound imaging and euthanasia, their bladders were collected and histopathological, immunohistochemical tests (ki67 e c-Myc) and Western Blotting (PI3K, mTOR, and p-4E-BP) were performed. RESULTS: Histopathological tests showed 66.67%, 62.5% and 37.5% incidence of BC in animals treated with BCG, NTZ, and NTZ + BCG, respectively. Nuclear positivity for ki-67 in BC animals were 12.4% (IC 10.1-14.6%), 13.2% (IC 10.5-15.9%) and 8.8% (IC 6.0-11.6%) in BCG, NTZ and NTZ + BCG group, respectively (p = 0.063). Between animals with carcinoma, c-Myc strong positive was 40.10% in NTZ, 32.2% in BCG and 19.90% in the NTZ + BCG group (p < 0.001). Blotting has shown mTOR (p = 0.0473) and PI3K inhibition (p = 0.0349) in the presence of BCG, added to 4-EBP inhibition in the presence of NTZ (p = 0.0004). CONCLUSIONS: Results show the possible synergy between the gold standard BC treatment BCG and NTZ, in which multiple targets inhibition such as c-Myc and downstream mTOR, p-4E-BP and PI3K might play a role.

Inhibition of urinary bladder cancer cell proliferation by silibinin.

Silibinin, a natural compound extracted from milk thistle, has demonstrated antitumor properties in urinary bladder cancer cells; however, the role of TP53 gene in these effects is unclear. In order to better understand the molecular and antiproliferative mechanisms of this compound, urinary bladder cancer cells with different TP53 gene status, RT4 (low-grade tumor, wild TP53 gene), 5637 (high-grade tumor, Grade 2, mutated TP53 gene), and T24 (high-grade tumor, Grade 3, mutated TP53 gene) were treated with several concentrations of silibinin (1, 5, 10, 50, 100, and 150 µM). Cytotoxicity, prooxidant effect, morphological changes, cell migration, cell cycle progression, global methylation profile, and relative expression of HOXB3, c-MYC, PLK1, SMAD4, SRC, HAT, HDAC, and RASSF1A genes were evaluated. The silibinin presented cytotoxic and prooxidant effects in the three cell lines. In mutated TP53 cells, significant interference in cell migration and cell cycle arrest at the G2/M phase was observed. Additionally, silibinin induced global DNA hypomethylation in the highest grade tumor cells. For wild-type TP53 cells, a sub-G1 apoptotic population was present. Furthermore, there was modulation of gene expression responsible for cell growth (SMAD and c-MYC), migration (SRC), cell cycle kinetics (PLK1), angiogenesis (HOXB3), and of genes associated with epigenetic events such as DNA acetylation (HAT) and deacetylation (HDAC). In conclusion, the silibinin inhibited the urinary bladder tumor cell proliferation independently of TP53 status; however, cell cycle effects, gene expression changes, and alteration of cell migration are dependent on TP53 status. © 2020 Wiley Periodicals, Inc.

Quinovic acid glycosides purified fraction from Uncaria tomentosa induces cell death by apoptosis in the T24 human bladder cancer cell line.

Bladder cancer is the second most prevalent malignancy in the genitourinary tract and remains a therapeutic challenge. In the search for new treatments, researchers have attempted to find compounds with low toxicity. With this goal in mind, Uncaria tomentosa is noteworthy because the bark and root of this species are widely used in traditional medicine and in adjuvant therapy for the treatment of numerous diseases. The objective of this study was to investigate the antitumor effect of one purified bioactive fraction of U.tomentosa bark on cell proliferation in two human bladder cancer cell lines, T24 and RT4. Quinovic acid glycosides purified fraction (QAPF) of U.tomentosa decreased the growth and viability of both T24 and RT4 cell lines. In T24 cells, QAPF induced apoptosis by activating caspase-3 and NF-κB. Further study showed that this fraction does not induce cell cycle arrest and does not alter PTEN and ERK levels. In conclusion, we demonstrated that QAPF of U.tomentosa has a potent inhibitory effect on the growth of human bladder cancer cell lines by inducing apoptosis through modulation of NF-κB, and we suggest that QAPF may become a potential therapeutic agent for the prevention and/or treatment of this cancer.

Chromosomal imbalances in successive moments of human bladder urothelial carcinoma.

OBJECTIVE: To understand developmental characteristics of urinary bladder carcinomas (UBC) by evaluating genomic alterations and p53 protein expression in primary tumors, their recurrences, and in the morphologically normal urothelium of UBC patients. METHODS: Tumors and their respective recurrences, six low-grade and five high-grade cases, provided 19 samples that were submitted to laser microdissection capture followed by high resolution comparative genomic hybridization (HR-CGH). HR-CGH profiles went through two different analyses--all tumors combined or classified according to their respective histologic grades. In a supplementary analysis, 124 primary urothelial tumors, their recurrences, and normal urothelium biopsied during the period between tumor surgical resection and recurrence, were submitted to immunohistochemical analyses of the p53 protein. During the follow-up of at least 21 patients, urinary bladder washes citologically negative for neoplastic cells were submitted to fluorescence in situ hybridization (FISH) to detect copy number alterations in centromeres 7, 17, and 9p21 region. RESULTS AND CONCLUSIONS: HR-CGH indicated high frequencies (80%) of gains in 11p12 and losses in 16p12, in line with suggestions that these chromosome regions contain genes critical for urinary bladder carcinogenesis. Within a same patient, tumors and their respective recurrences showed common genomic losses and gains, which implies that the genomic profile acquired by primary tumors was relatively stable. There were exclusive genomic alterations in low and in high grade tumors. Genes mapped in these regions should be investigated on their involvement in the urinary bladder carcinogenesis. Successive tumors from same patient did not present similar levels of protein p53 expression; however, when cases were grouped according to tumor histologic grades, p53 expression was directly proportional to tumor grades. Biopsies taken during the follow-up of patients with history of previously resected UBC revealed that 5/15 patients with no histologic alterations had more than 25% of urothelial cells expressing the p53 protein, suggesting that the apparently normal urothelium was genomically unstable. No numerical alterations of the chromosomes 7, 17, and 9p21 region were found by FISH during the periods "free-of-neoplasia." Our data are informative for further studies to better understand urinary bladder urothelial carcinogenesis.

Cistoprostatectomía radical laparoscópica con confección extracorpórea de neovejiga ortotópica: comunicación preliminar / Laparoscopic radical cystoprostatectomy and continent orthotopic neobladder performed extracorporeally: preliminary experience

Objetivo: Presentamos un abordaje quirúrgico para el manejo de pacientes con carcinoma vesical en candidatos a cirugía radical, buscando reunir los beneficios de la cirugía abierta tradicional, con las ventajas de la técnica laparoscópica. Material y Método: Se presenta una serie inicial de 5 pacientes de sexo masculino, llevados a cistoprostatectomía radical laparoscópica (CPRL) con confección extracorpórea de neovejiga ortotópica entre junio y septiembre de 2002. La edad promedio fue 50.6 años (rango: 29 a 70 años). La indicación quirúrgica fue carcinoma vesical infiltrante en cuatro de los cinco pacientes, uno de ellos previamente tratado con radioterapia. La técnica quirúrgica se presenta en 3 pasos. En el primero se realiza la linfadenectomía pelviana bilateral y la cistoprostatectomía radical (CPR) por laparoscopía; la segunda parte esta representada por la confección de la neovejiga, realizada completamente extracorpórea, y el tercer paso es la anastomosis uretra-neovejiga, desarrollada con técnica laparoscópica. Resultados: La técnica fue reproducida en los 5 pacientes. El tiempo operatorio promedio fue de 5.4 horas (rango: 4.5-7 horas). El sangrado promedio fue 410 ml (rango: 200-800 ml). Ninguno de los pacientes requirió transfusión sanguínea. El tiempo de hospitalización total varió entre 5 y 12 días, con un promedio de 6.8 días. Conclusión: La CPR puede realizarse completamente con técnica laparoscópica cuando se dispone de un adecuado entrenamiento laparoscópico. Con la técnica combinada que desarrollamos, la parte más complicada de la cirugía, representada por la elaboración de la neovejiga y el neoimplante ureteral, se realiza de forma convencional, haciendo que esta técnica sea totalmente reproducible.

Objective: We present our preliminary experience with laparoscopic radical cystoprostatectomy and continent orthotopic neobladder performed extracorporeally. We believe that this surgical approach combines the advantages of minimally invasive laparoscopy with the speed and safety of open surgery. Material and Method: Between June and September 2002, 5 male patients underwent to laparoscopic radical cystoprostatectomy and continent orthotopic neobladder performed extracorporeally. Average age was 50.6 years (range: 29-70). The operative indication was muscle-invasive carcinoma of the urinary bladder in 4 patients. One of these 4 patients was previously treated with radiotherapy. Our technique has 3 steps. Cystoprostatectomy and pelvic lymph node dissections are performed laparoscopically using five ports by a transperitoneal approach. We remove the surgical specimens through a 5 cm infraumbilical incision and through the same incision an ileal loop is extracted from the abdominal cavity, isolated, detubularized and neobladder is reconfigured. Finally, urethro-neobladder anastomosis is formed with laparoscopic technique. Results: Surgical technique was reproduced in all 5 patients. Mean operative time was 5.4 hours (range: 4.5-7). Mean blood loss was 410 ml (range: 200-800) and not transfusion was indicated. Mean hospital stay was 6.8 days (range: 5-12). Conclusion: Laparoscopic radical cystectomy is feasible when surgical team has experience in laparoscopic surgery. The most technically demanding steps of the procedure are neobladder confection and ureteral neoimplante, which are performed extracorporeally. We believe that with our combined approach, this technique is completely reproducible.