ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effect of combined use of oncolytic virus and the chemotherapeutic agents mitomycin (MMC) in growth inhibition of human bladder cancer cell line T-24 in vitro.</p><p><b>METHODS</b>Human bladder cancer cell line T-24 was infected with oncolytic virus (ONYX-015) of different multiplicity of infection, or treated with MMC in addition to ONYX-015. The changes in the cell growth, morphology, and apoptosis of cultured T-24 cells were observed by means of cell counting and fluorescence microscopy after the treatments. The effects of the treatment protocols were also tested in nude mouse model of implanted subcutaneous tumor.</p><p><b>RESULTS</b>Combined use of ONYX-015 and MMC produced substantially stronger cytotoxic effect against T-24 cells than exclusive use of ONYX-015. In in vivo experiments, combination of oncolytic virus and MMC resulted in much more significant tumor growth inhibition than either of the agents used alone. Obvious T-24 cell apoptosis could be observed in response to combined ONYX-105 and MMC treatment and exclusive ONYX-105 treatment.</p><p><b>CONCLUSIONS</b>ONYX-015 combined with MMC can produce significant cytotoxicity against T-24 cells and enhance therapeutic efficacy against bladder carcinoma.</p>
Subject(s)
Animals , Female , Mice , Antibiotics, Antineoplastic , Pharmacology , Therapeutic Uses , Apoptosis , Physiology , Cell Line, Tumor , Cell Proliferation , Cell Survival , Combined Modality Therapy , Mice, Inbred BALB C , Mice, Nude , Mitomycin , Pharmacology , Therapeutic Uses , Oncolytic Virotherapy , Methods , Oncolytic Viruses , Physiology , Urinary Bladder Neoplasms , Pathology , Therapeutics , Virology , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To label a human bladder cancer cell line and establish a novel human bladder cancer mouse model.</p><p><b>METHODS</b>T-24 cells, a human bladder transitional cell carcinoma cell line, were transfected with GFP plasmid to screen stable GFP-expressing clones. The latter were implanted into the wall of the bladder or the subcutaneous tissue of the neck of nude mice. The growth, invasion, and metastasis of the implanted tumor were observed and evaluated with whole-body optical imaging system. The findings were compared with those of HE staining on routine paraffin sections.</p><p><b>RESULTS</b>GFP-labeled tumor cells displayed green fluorescence under fluorescent microscopy and showed stable GFP expression in vitro and in vivo. One week after in situ transplantation of 5 x 10(5) T24 cells, the new bladder cancer was observed and evaluated under whole-body optical imaging system. Two weeks later, the new bladder tumor could be palpated, and 4 weeks later, metastasis to regional drainage lymph nodes in the pelvic and retroperitoneal lymph nodes occurred. The growth and metastasis of the implant bladder tumor were easily observed and accurately evaluated by fluorescent microscope.</p><p><b>CONCLUSION</b>GFP-labeled tumor cells display green fluorescence under fluorescent microscopy and show stable GFP expression. GFP-labeled T-24 cells and the novel human bladder cancer model described hereby provide a simple and reliable means for studying human bladder cancer in vivo.</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Carcinoma, Transitional Cell , Metabolism , Pathology , Diagnostic Imaging , Disease Models, Animal , Green Fluorescent Proteins , Genetics , Indicators and Reagents , Mice, Inbred BALB C , Mice, Nude , Microscopy, Fluorescence , Neoplasm Transplantation , Urinary Bladder Neoplasms , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the feasibility and efficacy of cytocine deaminase-thymidine kinase (CD-TK) fusion double suicide gene therapy using adenovirus mediated CD-TK gene and green fluorescent rotein (GFP) gene combined with ganciclovir(GCV) or 5-flourocytosine(5-FC) in a murine subcutaneous bladder carcinoma model.</p><p><b>METHODS</b>A replication defective adenovirus vector containing CD-TK gene was used. Subcutaneous tumors were established in syngenic C57BL/6 female mice with 1 x 10(6) Mb49 cells. Intratumoral injection of AdCD-TK (1.58 x 10(8) PFU, qd x days) in combination with GCV (40 mg.kg(-1).d(-1), ip, qd x 10 days) or 5-FC (400 mg.kg(-1).d(-1), ip, qd x 10 days) was administered in vivo for the determination of treatment efficacy in separate controlled experiments.</p><p><b>RESULTS</b>In vivo experiments demonstrated that the mean volume of tumor in the group of AdCD-TK/GCV(326.58+/-109.56 mm(3)), AdCD-TK/5-FC (235.33+/-62.94 mm(3)) and AdCD-TK/(GCV+5-FC) (23.58+/-6.78 mm(3)) was reduced significantly compared with that of control group (993.51+/-158.32 mm(3)) (P=0.00), the mean volume of tumor in the group of AdCD-TK/(GCV+5-FC) was significantly less than that in the group of AdCD-TK/GCV or AdCD-TK/5-FC (P=0.04). Tumor necrosis was revealed by histomorphology compared with control animals.</p><p><b>CONCLUSIONS</b>Adenovirus mediated CD-TK double suicide gene combining with GCV or 5-FC could provide an effective therapy in an experimental murine bladder carcinoma by significantly inhibiting tumor growth. The treatment efficacy of AdCD-TK combining GCV and 5-FC was superior to that of AdCD-TK combining GCV or AdCD-TK combining 5-FC.</p>
Subject(s)
Animals , Female , Mice , Adenoviridae , Genetics , Cell Line , Cell Line, Tumor , Cytosine Deaminase , Genetics , Metabolism , Defective Viruses , Genetics , Flucytosine , Pharmacology , Therapeutic Uses , Ganciclovir , Pharmacology , Therapeutic Uses , Genes, Transgenic, Suicide , Genetics , Genetic Therapy , Methods , Genetic Vectors , Mice, Inbred C57BL , Neoplasm Transplantation , Thymidine Kinase , Genetics , Metabolism , Treatment Outcome , Urinary Bladder Neoplasms , Pathology , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of apoptosis gene PDCD5 in tissues of normal human kidney, renal clear cell carcinoma, normal bladder and bladder carcinoma, and explore the role of PDCD5 gene in renal clear cell carcinoma and bladder carcinoma.</p><p><b>METHODS</b>Indirect immunohistochemistry was employed to detect PDCD5 expression in 63 kidney specimens and 42 bladder specimens. Positive expression rates and intensity of PDCD5 protein expression in the kidney tissue were investigated microscopically and by computerized image analysis. Positive expression rate in the bladder tissue was investigated by microscopic observation.</p><p><b>RESULTS</b>The results of immunohistochemical staining showed PDCD5 protein overexpression in the renal tubule of normal human kidney tissues and downregulation with the stage increase of renal clear cell carcinoma. PDCD5 protein expression showed statistical significance in tissues of normal kidney and renal clear cell carcinoma in all stages. No obvious PDCD5 expression was detected in the tissues of normal human bladder and bladder carcinoma.</p><p><b>CONCLUSION</b>PDCD5 is an important apoptosis-regulating factor in the occurrence of renal clear cell carcinoma, and its expression is extremely low in tissues of normal human bladder and bladder carcinoma.</p>