Resistance to anticancer chemotherapeutic drugs remains a major obstacle in cancer chemotherapy. A variety of mechanisms responsible for drug resistance has been posed. Mdr gene overexpression and detoxification by glutathione are believed to be involved in such mechanisms. Recently, we established two low-dose cisplatin-resistant human bladder cancer cell lines, T24RO.5 and T24R1, which showed resistance at O.5 hg/ml and 1 hg/ml of cisplatin, respectively. The resistance of T24RO.5 and T24R1 cells to cisplatin were 9.4 and 9.37 fold compared to that of the parental T24 cells In this study, we investigated the total glutathione content and p-glycoprotein expression, a mdr gene product, in parent and resistant cell lines to elucidate the drug resistance mechanism to cisplatin. Glutathione content was measured by biochemical method. P-glycoprotein expression was measured by flowcytometry using monoclonal antibody to p-glycoprotein. Glutathione content and p-glycoprotein expression were not different between parental and all resistant cell lines. These results suggest that mdr gene and glutathione do not play a role in cisplatin resistance mechanism in these low-dose cisplatin-resistant cell lines. Further work will be necessary to determine the mechanism of drug resistance in this model.
Cell Line* ; Cisplatin* ; Drug Resistance ; Drug Therapy ; Genes, MDR* ; Glutathione* ; Humans ; Metabolism* ; P-Glycoprotein ; Parents ; Urinary Bladder Neoplasms ; Urinary Bladder*

OBJECTIVETo study the clinical and pathologic characteristics of small cell neuroendocrine carcinoma of urinary tract.

METHODSAll cases of urinary tract carcinoma encountered in the General Hospital of People Liberation Army during the period from 1999 to 2010 were retrospectively reviewed. The clinicopathologic data of small cell neuroendocrine carcinomas were further analyzed, with literature review.

RESULTSA total of 16 cases of small cell neuroendocrine carcinoma were identified, including 10 from urinary bladder, 2 from ureter, 3 from renal pelvis, and 1 multifocal tumor involving renal pelvis and ureter. There were altogether 8 males and 8 females. The median age of the patients was 63 years (range = 24 to 79 years). Gross hematuria (11 cases) represented the main presenting symptom. Four patients had flank pain and 4 had urinary irritation symptoms. Seven patients underwent radical cystectomy. Six other patients underwent radical nephroureterectomy, 1 partial cystectomy, 1 TURBT and the remaining case biopsy only. The size of the tumor ranged from 0.8 to 8.0 cm (median = 4.5 cm). Histologically, 15 cases represented mixed small cell neuroendocrine carcinoma (with 13 mixed with transitional cell carcinoma and 2 with adenocarcinoma). Immunohistochemical study showed positive staining for neuroendocrine markers. On presentation, 1 patient was in stage pT1, 7 in stage pT2, 6 in stage pT3, 2 in stage pT4. Six patients died of the disease after operation. The overall survival was 25 months and the 5-year survival rate was 32.4%.

CONCLUSIONSSmall cell neuroendocrine carcinoma of urinary bladder is a highly malignant disease and associated with poor prognosis. The diagnosis relies on detailed histologic examination. Early diagnosis, when coupled with cystectomy or nephroureterectomy and adjuvant chemotherapy, represents the mainstay of management.

Adult ; Aged ; CD56 Antigen ; metabolism ; Carcinoma, Neuroendocrine ; drug therapy ; metabolism ; pathology ; surgery ; Carcinoma, Small Cell ; drug therapy ; metabolism ; pathology ; surgery ; Chemotherapy, Adjuvant ; Cystectomy ; Female ; Follow-Up Studies ; Humans ; Keratins ; metabolism ; Kidney Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Nephrectomy ; Phosphopyruvate Hydratase ; metabolism ; Retrospective Studies ; Survival Rate ; Synaptophysin ; metabolism ; Ureteral Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Urinary Bladder Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Urologic Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Young Adult

Although an immune response to bacillus Calmette Guerin (BCG) has often been associated with antitumor activity, the action mechanism(s) of intravesical BCG therapy for prophylaxis and treatment of superficial bladder cancer is not clearly understood. In an attempt to evaluate the roles of tumor necrosis factor (TNF)-alpha and lymphotoxin (LT) in the antitumor activity, TNF-alpha productivities by peripheral blood monocytes, serum levels of TNF-alpha, and LT productivities by peripheral blood lymphocytes were studied in superficial bladder cancer patients after six intravesical administrations of BCG. TNF-alpha productivities by peritoneal macrophages of guinea pigs were also studied after six intravesical administrations of BCG. The maximum TNF-alpha productivities by peripheral blood monocytes of superficial bladder cancer patients were seen after the fourth week of administration of BCG, and the serum TNF-alpha levels were also slightly increased after intravesical BCG administration in the superficial bladder cancer patients. LT productivities by peripheral blood lymphocytes of superficial bladder cancer patients were significantly enhanced and the maximum LT productivity was also seen after the third or fifth BCG administration. TNF-alpha productivities by peritoneal macrophages of guinea pigs were significantly enhanced and the maximum TNF-alpha productivity was seen after the second or third BCG administration. Our data might suggest that six consecutive intravesical BCG administrations could induce the increased productions of TNF-alpha and LT, which might play an important role in the antitumor activity in superficial bladder cancer.Although an immune response to bacillus Calmette Guerin (BCG) has often been associated with antitumor activity, the action mechanism(s) of intravesical BCG therapy for prophylaxis and treatment of superficial bladder cancer is not clearly understood. In an attempt to evaluate the roles of tumor necrosis factor (TNF)-alpha and lymphotoxin (LT) in the antitumor activity, TNF-alpha productivities by peripheral blood monocytes, serum levels of TNF-alpha, and LT productivities by peripheral blood lymphocytes were studied in superficial bladder cancer patients after six intravesical administrations of BCG. TNF-alpha productivities by peritoneal macrophages of guinea pigs were also studied after six intravesical administrations of BCG. The maximum TNF-alpha productivities by peripheral blood monocytes of superficial bladder cancer patients were seen after the fourth week of administration of BCG, and the serum TNF-alpha levels were also slightly increased after intravesical BCG administration in the superficial bladder cancer patients. LT productivities by peripheral blood lymphocytes of superficial bladder cancer patients were significantly enhanced and the maximum LT productivity was also seen after the third or fifth BCG administration. TNF-alpha productivities by peritoneal macrophages of guinea pigs were significantly enhanced and the maximum TNF-alpha productivity was seen after the second or third BCG administration. Our data might suggest that six consecutive intravesical BCG administrations could induce the increased productions of TNF-alpha and LT, which might play an important role in the antitumor activity in superficial bladder cancer.
Administration, Intravesical ; Animal ; Bladder Neoplasms/*metabolism/*therapy ; Female ; Guinea Pigs ; Human ; Mycobacterium bovis/*physiology ; Prospective Studies ; Support, Non-U.S. Gov't ; Tumor Necrosis Factor/*biosynthesis

BACKGROUNDSuperficial urothelial carcinoma (SUC) of the bladder is a common urinary tract tumor in China. There is a high recurrence rate of this tumor even after surgery and intravesical instillation. Previous reports have described a suppression of the immune system in cancer patients. Dendritic cells (DCs) play a pivotal role in the induction of an effective antitumor immune response. The aim of this study was to investigate the effects of surgery and epirubicin intravesical chemotherapy (IC) on peripheral blood DCs in subsets of patients with bladder SUC.

METHODSA total of 66 SUC patients and 38 healthy controls were enrolled in this study. All the patients had undergone transurethral resection (TUR) of their cancer and adjunctive IC after tumor removal. The patients were divided into a non-recurrence group (n = 40) and a recurrence group (n = 26) based on the presence or absence of tumor recurrence. Blood samples were taken preoperatively (PreOP), on postoperative days (POD) 1 and 7, and at postoperative month (POM) 3. Flow cytometric analysis was used for the determination and quantitation of the surface markers CD80 and CD86 in circulating DC subsets.

RESULTSThe preoperative percentages of myeloid dendritic cells (mDCs) and expression of CD80 and CD86 were impaired in SUC patients compared to healthy controls (P < 0.05). The percentages of mDCs and these surface markers decreased significantly on POD 1 and increased on POD 7, remaining higher than the preoperative values in POM 3 (P < 0.05). The percentages of mDCs, and CD80 and CD86 in the non-recurrence group on PreOP, POD 7, and POM 3 were higher than those in recurrence group.

CONCLUSIONSSurgical removal of SUC and adjunctive IC were associated with improved circulating mDC counts and function. Persistent depression of mDC counts and function after treatment in recurrence patients indicated lower antitumor immunity that may lead to tumor recurrence.

Adult ; Aged ; China ; Dendritic Cells ; immunology ; metabolism ; Epirubicin ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Urinary Bladder Neoplasms ; drug therapy ; immunology ; surgery

BACKGROUND AND OBJECTIVEIntravesical administration of Bacillus Calmette-Guerin (BCG) after transurethral resection is by far the most effective local therapy for superficial bladder cancer, the fifth most common cancer in the world. However, approximately one-third of patients fail to respond and most patients eventually relapse. In addition, there are pronounced side effects of BCG therapy, such as BCG sepsis and a high frequency of BCG-induced cystitis. This study established a novel immunotherapy through immobilization of streptavidin-tagged human IL-2 (SA-hIL-2) on the biotinylated mucosal surface of bladder wall.

METHODSA mouse orthotopic model of MB49 bladder cancer was established by perfusing MB49 cells into mouse bladders. The SA-hIL-2 fusion protein was immobilized on the biotinylated mucosal surface of the bladder wall. Treatment began on day 1 after MB49 implantation, once every 3 days for 6 times. Immunohistochemical assay was performed to assess the persistence of SA-hIL-2 immobilized on the biotinylated mucosal surface of the bladder wall. The mice were monitored for tumor growth and survival. On day 60 after MB49 implantation, the SA-hIL-2-cured mice, which were found to have no hematuria or palpable tumors, were challenged with wild-type MB49 cells implanted into the pretreated bladder and monitored for survival.

RESULTSSA-hIL-2 could be immobilized efficiently and durably on the bladder mucosal surface as long as 7 days. On day 60 after MB49 implantation, 9 out of 20 SA-hIL-2-treated mice survived, but all mice in PBS control group died. More importantly, 5 out of 9 tumor-free mice in the SA-hIL-2 group were protected against a second intravesical wild-type MB49 tumor challenge.

CONCLUSIONSSA-hIL-2 fusion protein could significantly inhibit tumor growth and extend the survival time in the orthotopic model of MB49 bladder cancer.

Animals ; Biotinylation ; Cell Line, Tumor ; Female ; Immobilized Proteins ; metabolism ; therapeutic use ; Immunotherapy ; methods ; Interleukin-2 ; metabolism ; therapeutic use ; Mice ; Mice, Inbred C57BL ; Mucous Membrane ; metabolism ; Neoplasm Transplantation ; Receptors, Interleukin-2 ; metabolism ; Recombinant Fusion Proteins ; metabolism ; therapeutic use ; Streptavidin ; metabolism ; therapeutic use ; Urinary Bladder ; pathology ; Urinary Bladder Neoplasms ; immunology ; therapy

OBJECTIVETo evaluate the apoptosis-inducing effect of intravesical instillation of pirarubicin (THP) on bladder cancer and normal bladder tissue, and explore the mechanism of such treatment for preventing recurrence of non-muscle invasive bladder cancer.

METHODSForty patients with primary non-muscle invasive bladder cancer were treated in our hospital from January 2006 to October 2008. The patients were divided into three groups, including 15 cases treated by intravesical instillation of THP (30 mg/50 ml, 0.5 hours) at 1 hour, 15 cases at 24 hours before transuretheral resection of bladder tumor (TUR-BT), and 10 cases in control group who received TUR-BT only. The THP uptake of the tumor and normal bladder tissues was observed by fluorescence microscopy, the apoptosis was assessed with TUNEL staining, and immunohistochemistry was used to detect the expression of bcl-2 and bax.

RESULTSSeldom fluorescence of THP in normal bladder tissues and some diffuse fluorescence reaching the muscular layer in the tumor area were observed after THP instillation at 1 hour before TUR-BT. The fluorescence of THP could still be observed in the tumor tissues at 24 hours after THP instillation. The apoptosis indexes (AI) of tumors in the chemotherapy groups were significantly higher than that in the control group (P < 0.01). There was a correlation between bcl-2/bax ratio and AI.

CONCLUSIONPirarubicin can be uptaken by bladder cancer tissue selectively and induces apoptosis of tumor cells. The changes of expression of bcl-2 and bax and decreasing of the bcl-2/bax ratio may be an important mechanism of action of the intravesical instillation of THP for preventing recurrence of non-muscle invasive bladder cancer.

Administration, Intravesical ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; administration & dosage ; metabolism ; therapeutic use ; Apoptosis ; drug effects ; Cystectomy ; Doxorubicin ; administration & dosage ; analogs & derivatives ; metabolism ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Preoperative Care ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Urinary Bladder ; metabolism ; pathology ; Urinary Bladder Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE: To explore the influences of andrographolide (Andro) on bladder cancer cell lines and a tumor xenograft mouse model bearing 5637 cells. METHODS: For in vitro experiments, T24 cells were stimulated with Andro (0-40 µmol/L) and 5637 cells were stimulated with Andro (0 to 80 µmol/L). Cell growth, migration, and infiltration were assessed using cell counting kit-8, colony formation, wound healing, and transwell assays. Apoptosis rate was examined using flow cytometry. In in vivo study, the antitumor effect of Andro (10 mg/kg) was evaluated by 5637 tumor-bearing mice, and levels of nuclear factor κ B (NF- κ B) and phosphoinositide 3-kinase/AKT related-proteins were determined by immunoblotting. RESULTS: Andro suppressed growth, migration, and infiltraion of bladder cancer cells (P⩽0.05 or P⩽0.01). Additionally, Andro induced intrinsic mitochondria-dependent apoptosis in bladder cancer cell lines. Furthermore, Andro inhibited bladder cancer growth in mice (P⩽0.01). The expression of p65, p-AKT were suppressed by Andro treatment in vitro and in vivo (P⩽0.05 or P⩽0.01). CONCLUSIONS Andrographolide inhibits proliferation and promotes apoptosis in bladder cancer cells by interfering with NF- κ B and PI3K/AKT signaling in vitro and in vivo.
Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes/therapeutic use* ; Humans ; Mice ; NF-kappa B/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Urinary Bladder Neoplasms/drug therapy*

Bmi1 is a member of the polycomb group family of proteins, and it drives the carcinogenesis of various cancers and governs the self-renewal of multiple types of stem cells. However, its role in the initiation and progression of bladder cancer is not clearly known. The present study aimed to investigate the function of Bmi1 in the development of bladder cancer. Bmi1 expression was detected in human bladder cancer tissues and their adjacent normal tissues (n=10) by immunohistochemistry, qRT-PCR and Western blotting, respectively. Bmi1 small interference RNA (siRNA) was synthesized and transfected into human bladder carcinoma cells (EJ) by lipofectamine 2000. The Bmil expression at mRNA and protein levels was measured in EJ cells transfected with Bmil siRNA (0, 80, 160 nmol/L) by qRT-PCR and Western blotting, respectively. Cell viability and Ki67 expression (a marker of cell proliferation) were determined in Bmi1 siRNA-transfected cells by CCK-8 assay and qRT-PCR, respectively. Cell cycle of transfected cells was flow-cytometrically determined. Immunofluorescence and Western blotting were used to detect the expression levels of cell cycle-associated proteins cyclin D1 and cyclin E in the cells. Pro-apoptotic proteins Bax and caspase 3 and anti-apoptotic protein Bcl-2 were detected by Western blotting as well. Additionally, xenograft tumor models were established by inoculation of EJ cells (infected with Bmil shRNA/pLKO.1 lentivirus or not) into nude mice. The tumor volumes were measured every other day for 14 days. The results showed that the Bmil expression was significantly increased in bladder tumor tissues when compared with that in normal tissues (P<0.05). Perturbation of Bmi1 expression by using siRNA could significantly inhibit the proliferation of EJ cells (P<0.05). Bmi1 siRNA-transfected EJ cells were accumulated in G1 phase and the expression levels of cyclin D1 and cyclin E were down-regulated. Bax and caspase-3 expression levels were significantly increased and Bcl-2 levels decreased after Bmi1 knockdown. Tumor volume was conspicuously reduced in mice injected with EJ cells with Bmi1 knockdown. Our findings indicate that Bmi1 is a potential driver oncogene of bladder cancer and it may become a potential treatment target for human bladder cancer.
Animals ; Apoptosis ; genetics ; Carcinogenesis ; genetics ; metabolism ; pathology ; Carcinoma ; genetics ; metabolism ; pathology ; therapy ; Caspase 3 ; genetics ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; antagonists & inhibitors ; genetics ; metabolism ; Cyclin E ; antagonists & inhibitors ; genetics ; metabolism ; G1 Phase Cell Cycle Checkpoints ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Injections, Intralesional ; Ki-67 Antigen ; genetics ; metabolism ; Mice ; Mice, Nude ; Polycomb Repressive Complex 1 ; antagonists & inhibitors ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; genetics ; metabolism ; RNA, Small Interfering ; administration & dosage ; genetics ; metabolism ; Signal Transduction ; Tumor Burden ; Urinary Bladder ; metabolism ; pathology ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology ; therapy ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; agonists ; genetics ; metabolism

No abstract available.
Aged ; Anti-Bacterial Agents/therapeutic use ; Bacteria/metabolism ; Catheter-Related Infections/diagnosis/drug therapy/*microbiology ; Color ; Equipment Design ; Escherichia coli Infections/diagnosis/drug therapy/*microbiology ; Female ; Humans ; Intestines/*microbiology ; Pigments, Biological/metabolism ; Treatment Outcome ; Tryptophan/metabolism ; Urinary Bladder Neoplasms/surgery ; Urinary Catheterization/adverse effects/*instrumentation ; *Urinary Catheters ; *Urinary Diversion ; Urinary Tract Infections/diagnosis/drug therapy/*microbiology ; Urine/chemistry/microbiology

OBJECTIVETo explore multi-drug resistance (MDR) of bladder cancer for the intravesical instillation.

METHODSUsing immunohistochemical staining, in 44-case human bladder cancer cells, the expressions of P-glycoprotein (P-gp), glutathione S-transferase (GST-pi) and topoisomerase (TOPO-II), were detected to find out the resistance to drugs.

RESULTSP-gp had a higher expression in 54.5% cases. GST-pi had no or a lower expression in 65.9% cases. TOPO-II had a higher expression in 29.5% but a lower expression in 65.9% cases.

CONCLUSIONDetecting the factors of MDR in bladder cancer cells could help to choose drugs for intravesical chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Administration, Intravesical ; Adult ; Aged ; DNA Topoisomerases, Type II ; analysis ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Glutathione Transferase ; analysis ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Urinary Bladder Neoplasms ; drug therapy ; metabolism