tetracycline and Urinary-Bladder-Neoplasms

Topics: Carcinoma; Cystoscopy; Fluorescence; History, 20th Century; Humans; Optical Imaging; Photosensitizing Agents; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms

Previous studies have suggested that EZH2 is up-regulated in bladder cancer tissues and identified it as a biomarker for poor prognosis. However, the biological functions of EZH2 in bladder cancer cells remain unknown. In this research, we discovered that EZH2 expression is irrelevant to the TNM stage and poor prognosis of bladder cancer patients. But suppression of EZH2 can slowdown the progression of bladder cancer cells. Moreover, we used the technology of synthetic biology to construct the tetracycline-controllable artificial microRNA-HOTAIR + EZH2, which can decrease the expression of HOTAIR and EZH2 in a doxycycline dosage-dependent manner. And we also found that HOTAIR expression was positively correlated with EZH2 expression. Tetracycline-controllable artificial microRNA-HOTAIR + EZH2 can inhibit the proliferation and migration of bladder cancer cells. Meanwhile, the apoptosis rate of bladder cancer cells was increased. Taken together, our research showed the cancer-promoting effects of EZH2 and created a novel method to rescue the development of bladder cancer cells.

Topics: Antagomirs; Apoptosis; Base Sequence; Cell Line, Tumor; Cell Movement; Cell Proliferation; Databases, Factual; Disease Progression; Enhancer of Zeste Homolog 2 Protein; Gene Expression Regulation, Neoplastic; Humans; Neoplasm Grading; RNA, Long Noncoding; RNA, Small Interfering; Tetracycline; Urinary Bladder Neoplasms

Small hairpin RNA (shRNA) can inhibit the malignant phenotypes of tumor cell through ribonucleic acid interference (RNAi). However, it is hardly to be regulated and it may induce few phenotypic changes. Here, we build a type of tetracycline (Tet)-inducible vectors which can achieve regulatable expression of shRNA in a time-dependent manner by using synthetic biology approach. In order to prove the effectiveness of this device, we chose hTERT and Bcl-2 as target genes and test the utility of the device on 5637 and T24 cell lines. The experiments show that the Tet-inducible small hairpin RNA can effectively suppress their target genes and generate anti-cancer effects on both 5637 and T24 cell lines. The device we build not only can inhibit proliferation but also can induce apoptosis and suppress migration of the bladder cancer cell lines 5637 and T24. The Tet-inducible small hairpin RNAs may provide a novel strategy for the treatment of human bladder cancer in the future.

Topics: Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Humans; Phenotype; Proto-Oncogene Proteins c-bcl-2; RNA, Small Interfering; Telomerase; Tetracycline; Urinary Bladder Neoplasms

Recent reports show that long non-coding RNAs (lncRNAs) are emerging as significant functional regulators in the development of tumors, including bladder cancer. Here, we found that CCAT2 was upregulated in bladder cancer tissues and cell lines. Through the statistical analyses, we also found that the high expression level of CCAT2 was positively correlated with histological grade and TNM stage of bladder cancer. Further experimental results revealed that knockdown of CCAT2 could decrease cell proliferation and migration as well as induce apoptosis in bladder cancer cells. Besides, using the post-transcriptional device of synthetic biology, we create the tetracycline-inducible double small hairpin RNAs (shRNAs) vector to control the expression level of CCAT2 which was induced by doxycycline in a dosage-dependent manner. In summary, our data indicated that CCAT2 may be an oncogene and a therapeutic target in bladder cancer. The expression of CCAT2 can be quantitatively controlled by the synthetic "tetracycline-on" switch system in bladder cancer in response to different concentrations of doxycycline to inhibit the development of bladder cancer cells.

Topics: Adult; Aged; Apoptosis; Disease Progression; Female; Gene Expression Regulation, Neoplastic; Genetic Therapy; Genetic Vectors; Humans; Male; Middle Aged; RNA, Long Noncoding; RNA, Small Interfering; Tetracycline; Urinary Bladder Neoplasms

Various studies have indicated that long non-coding RNAs (lncRNAs) play vital roles in the cancer development and progression. LncRNA hypoxia inducible factor 1alpha antisense RNA-2 (HIF1A-AS2) is upregulated in gastric carcinomas and knockdown of HIF1A-AS2 expression by siRNA could inhibit cell proliferation in vitro and tumorigenesis in vivo. Inspired by these observations, we hypothesized that HIF1A-AS2 possibly plays the analogous roles in bladder cancer. In our study, we first reported that HIF1A-AS2 was up-regulated in bladder cancer tissues and cells, and HIF1A-AS2 expression level in bladder cancer tissues is positively associated with advanced clinical pathologic grade and TNM phase. Cell proliferation inhibition, cell migration suppression and apoptosis induction were observed by silencing HIF1A-AS2 in bladder cancer T24 and 5637 cells. Overexpression of HIF1A-AS2 in SV-HUC-1 cells could promote cell proliferation, cell migration and anti-apoptosis. Besides, we utilized the emerging technology of medical synthetic biology to design tetracycline-inducible small hairpin RNA (shRNA) vector which specifically silenced HIF1A-AS2 in a dosage-dependent manner to inhibit the progression of human bladder cancer. In conclusion, our data suggested that HIF1A-AS2 plays oncogenic roles and can be used as a therapeutic target for treating human bladder cancer. Synthetic "tetracycline-on" switch system that quantitatively controlled the expression of HIF1A-AS2 in bladder cancer can inhibit the progression of bladder cancer cells in a dosage-dependent manner. Our findings provide new insights into the role of the lncRNA HIF1A-AS2 in the bladder cancer.

Topics: Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Neoplasm Invasiveness; Phenotype; RNA Interference; RNA, Long Noncoding; RNA, Small Interfering; RNAi Therapeutics; Tetracycline; Time Factors; Transfection; Up-Regulation; Urinary Bladder Neoplasms

Recent studies show that long non-coding RNAs (lncRNAs) may be significant functional regulators in tumor development, including bladder cancer. Here, we found that PVT1 was upregulated in bladder cancer tissues and cells. Further experiments revealed that PVT1 promoted cell proliferation and suppressed cell apoptosis. Furthermore we also used the emerging technology, synthetic biology, to create tetracycline-inducible small hairpin RNA (shRNA) vectors which silenced PVT1 in a dosage-dependent manner to inhibit the progression of bladder cancer. In conclusion, data suggest that PVT1 could be an oncogene and may be a therapeutic target in bladder cancer. Synthetic "tetracycline-on" switch system can be used to quantitatively control the expression of PVT1 in bladder cancer in response to different concentration of doxycycline to suppress the progression of bladder cancer.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Cell Line; Cell Line, Tumor; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Neoplasm Grading; Neoplasm Staging; Reverse Transcriptase Polymerase Chain Reaction; RNA, Long Noncoding; RNA, Small Interfering; Tetracycline; Up-Regulation; Urinary Bladder Neoplasms

Ribonucleic acid interference (RNAi) based on microRNA (miRNA) may provide efficient and safe therapeutic opportunities. However, natural microRNAs can not easily be regulated and usually cause few phenotypic changes. Using the engineering principles of synthetic biology, we provided a novel and standard platform for the generation of tetracycline (Tet)-inducible vectors that express artificial microRNAs in a dosage-dependent manner. The vector generates a Pol II promoter-mediated artificial microRNA which was flanked by ribozyme sequences. In order to prove the utility of this platform, we chose β-catenin and HIF-1α as the functional targets and used the bladder cancer cell lines 5637 and T24 as the test models. We found that the Tet-inducible artificial microRNAs can effectively silence the target genes and their downstream genes, and induce anti-cancer effects in the two bladder cancer cell lines. These devices can inhibit proliferation, induce apoptosis, and suppress migration of the bladder cancer cell lines 5637 and T24. The Tet-inducible synthetic artificial microRNAs may represent a kind of novel therapeutic strategies for treating human bladder cancer.

Topics: Apoptosis; beta Catenin; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; MicroRNAs; Phenotype; Promoter Regions, Genetic; Tetracycline; Urinary Bladder Neoplasms

A cross-talk between different receptor tyrosine kinases (RTKs) plays an important role in the pathogenesis of human cancers.. Both NIH-Met5 and T24-Met3 cell lines harboring an inducible human c-Met gene were established. C-Met-related RTKs were screened by RTK microarray analysis. The cross-talk of RTKs was demonstrated by Western blotting and confirmed by small interfering RNA (siRNA) silencing, followed by elucidation of the underlying mechanism. The impact of this cross-talk on biological function was demonstrated by Trans-well migration assay. Finally, the potential clinical importance was examined in a cohort of 65 cases of locally advanced and metastatic bladder cancer patients.. A positive association of Axl or platelet-derived growth factor receptor-alpha (PDGFR-α) with c-Met expression was demonstrated at translational level, and confirmed by specific siRNA knock-down. The transactivation of c-Met on Axl or PDGFR-α in vitro was through a ras- and Src-independent activation of mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK/ERK) pathway. In human bladder cancer, co-expression of these RTKs was associated with poor patient survival (p < 0.05), and overexpression of c-Met/Axl/PDGFR-α or c-Met alone showed the most significant correlation with poor survival (p < 0.01).. In addition to c-Met, the cross-talk with Axl and/or PDGFR-α also contributes to the progression of human bladder cancer. Evaluation of Axl and PDGFR-α expression status may identify a subset of c-Met-positive bladder cancer patients who may require co-targeting therapy.

Topics: Animals; Axl Receptor Tyrosine Kinase; Cell Line, Tumor; Cell Movement; Extracellular Signal-Regulated MAP Kinases; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Mice; NIH 3T3 Cells; Oncogene Protein p21(ras); Oncogene Protein pp60(v-src); Prognosis; Protein Binding; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-met; Receptor Protein-Tyrosine Kinases; Receptor, Platelet-Derived Growth Factor alpha; Signal Transduction; Survival Analysis; Tetracycline; Transcriptional Activation; Urinary Bladder Neoplasms

We have shown previously that wild type p53 can rapidly induce replicative senescence in EJ human bladder carcinoma cells lacking functional p53. A major effector of p53 functions is p21Waf1/Cip1/Sdi1, a potent cyclin-dependent kinase inhibitor. p21Waf1/Cip1/Sdi1 has been shown to be involved in both p53 dependent and independent control of cell proliferation, differentiation and death. To directly investigate the effects of p21Waf1/Cip1/Sdi1 in the p53 response observed in EJ tumor cells, we established p21Waf1/Cip1/Sdi1 inducible lines using the tetracycline-regulatable vector system. p21Waf1/Cip1/Sdi1 induction caused irreversible cell cycle arrest in both G1 and G2/M, and diminished Cdk2 kinase activity. In addition, p21Waf1/Cip1/Sdi1 induction led to morphological alterations characteristic of cells undergoing replicative senescence with morphological, biochemical and ultrastructural markers of the senescent phenotype. Furthermore, sustained p21Waf1/Cip1/Sdi1 induction sensitized EJ cells to apoptotic cell death induced by mitomycin C, a cross-linking DNA damaging agent. These findings support the function of p21Waf1/Cip1/Sdi1 as an inducer of replicative senescence and a major mediator of this phenomenon in response to p53. Moreover, our results imply that therapeutic intervention in human cancers might be aimed at sustained elevation of p21Waf1/Cip1/Sdi1 expression.

Topics: Apoptosis; beta-Galactosidase; Biomarkers; Carcinoma; CDC2-CDC28 Kinases; Cell Division; Cellular Senescence; Cross-Linking Reagents; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinases; Cyclins; G1 Phase; G2 Phase; Gene Expression Regulation, Neoplastic; Genetic Engineering; Humans; Mitomycin; Protein Serine-Threonine Kinases; Tetracycline; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

p53 is mutated in approximately 50% of human cancers, whereas mutations of the related p73 gene are rare. p73 can activate p53-responsive promoters and induce apoptosis when overexpressed in certain p53-deficient tumor cells. We show that p73 isoforms, p73alpha and p73beta, can each induce permanent growth arrest with markers of replicative senescence when overexpressed in a tetracycline-regulatable manner in human cancer cells lacking functional p53. Human homologue of mouse double minute 2 gene product (hMDM2), but not an NH(2)-terminal deletion mutant, coimmunoprecipitated with p73alpha or p73beta, and inhibited p73 transcriptional activity as with p53. In contrast to p53, ectopically expressed hemagglutinin (HA)-tagged p73 proteins were not stabilized by treatment with several DNA damaging agents. Furthermore, unlike normal p53, which increases in response to DNA damage due to enhanced protein stability in MCF7 cells, endogenous p73 protein levels were not increased in these cells under the same conditions. Thus, although p73 has an ability, comparable to that of p53, to suppress tumor cell growth in p53-deficient cells, p73 induction is regulated differently from p53. These findings suggest that the selective pressures for p53 rather than p73 inactivation in tumors may reflect their differential responses to stresses such as DNA damage, rather than their capacities to induce permanent growth arrest or apoptosis programs.

Topics: Animals; Breast Neoplasms; Cell Cycle; Cell Line; DNA Damage; DNA-Binding Proteins; Female; Genes, Tumor Suppressor; Humans; Lung Neoplasms; Mice; Neoplasm Proteins; Nuclear Proteins; Protein Isoforms; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-mdm2; Recombinant Proteins; Tetracycline; Transcription, Genetic; Transfection; Tumor Cells, Cultured; Tumor Protein p73; Tumor Suppressor Protein p53; Tumor Suppressor Proteins; Urinary Bladder Neoplasms

Topics: Carcinoma, Transitional Cell; Chemical Phenomena; Chemistry; Coloring Agents; Cystoscopy; Fluorescent Dyes; Hematoporphyrins; Humans; Methylene Blue; Tetracycline; Urinary Bladder Neoplasms

Positive results were obtained in epithelial concentrations when tetracycline was given to 50 patients with bladder tumors. The diagnosis of bladder tumors with epithelial concentration is especially helpful in patients with hematuria and pyuria. With this technique many epithelial cells radiate a bright yellow fluorescence.

Topics: Adult; Aged; Cytodiagnosis; Epithelial Cells; Epithelium; Female; Fluorescence; Hematuria; Humans; Male; Middle Aged; Pyuria; Staining and Labeling; Tetracycline; Urinary Bladder Neoplasms

Topics: Biopsy; Cystoscopy; Humans; Photochemotherapy; Photofluorography; Tetracycline; Urinary Bladder Neoplasms

The clinical, laboratory, and radiographic characteristics of polypoid cystitis in 2 dogs were similar to those commonly associated with neoplasms of the urinary bladder. Gross appearance of the polyps did not permit differentiation between inflammation and neoplasia; microscopic examination of excised tissue was required to establish a definitive diagnosis. Following surgical extirpation of the polyps, remission of clinical signs was induced by prolonged antibacterial therapy.

Topics: Ampicillin; Animals; Bacteriuria; Chloramphenicol; Cystitis; Dog Diseases; Dogs; Female; Gentamicins; Hematuria; Male; Polyps; Proteus; Radiography; Tetracycline; Urinary Bladder; Urinary Bladder Neoplasms

Topics: Carcinoma; Cystoscopy; Fluorometry; Humans; Papilloma; Tetracycline; Time Factors; Ultraviolet Rays; Urinary Bladder Neoplasms

Topics: Biopsy; Carcinoma, Squamous Cell; Cystoscopy; Fluorescence; Follow-Up Studies; Humans; Methods; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms; Urine

Topics: Abdomen, Acute; Aged; Blood Urea Nitrogen; Humans; Kidney Calculi; Kidney Diseases; Male; Middle Aged; Tetracycline; Uremia; Urinary Bladder Neoplasms

Topics: Cystoscopy; Fluorescence; Humans; Microscopy, Fluorescence; Staining and Labeling; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms

Topics: Acridines; Cystoscopes; Cystoscopy; Fluorescence; Humans; Methods; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms

Topics: Animals; Autoradiography; Female; Rats; Tetracycline; Tritium; Urinary Bladder Diseases; Urinary Bladder Neoplasms

Topics: Cystoscopy; Fluorescence; Humans; Tetracycline; Urinary Bladder Neoplasms

Topics: Fluorescence; Humans; Male; Prostatic Neoplasms; Tetracycline; Urinary Bladder Neoplasms

Topics: Anti-Bacterial Agents; Humans; Methacycline; Microscopy, Fluorescence; Spectrophotometry; Staining and Labeling; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms

Topics: Biomedical Research; Cystitis; Fluorescence; Humans; Kidney Neoplasms; Male; Neoplasms; Prostatic Neoplasms; Tetracycline; Urinary Bladder Neoplasms; Urinary Diversion; Urine

Topics: Adenocarcinoma; Carcinoma; Carcinoma, Squamous Cell; Cystoscopy; Fluorescence; Humans; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms

Topics: Anti-Bacterial Agents; Candidiasis; Chemical and Drug Induced Liver Injury; Chloramphenicol; Diagnosis, Differential; Erythromycin; Halothane; Hepatitis; Hydrocortisone; Hypertension; Kidney Diseases; Metaraminol; Novobiocin; Pathology; Pyelonephritis; Surgical Procedures, Operative; Surgical Wound Infection; Tetracycline; Toxicology; Tracheotomy; Urinary Bladder Neoplasms; Urinary Diversion

Topics: Anti-Bacterial Agents; Fluorescence; Geriatrics; Humans; Neoplasms; Pathology; Surgical Procedures, Operative; Tetracycline; Ultraviolet Rays; Urinary Bladder Neoplasms