ABSTRACT
OBJECTIVE: Investigating whether extracellular factors are possible actors in tumoral progression in bladder carcinoma. METHODS: RT112/G2 bladder tumour cells were grown in presence of TGFbeta and analysed by immunological and cDNA microarray techniques. RESULTS: TGFbeta inhibited cell proliferation, reduced TNFalpha- and IFNgamma-induced apoptosis by decreasing TNFalpha-RI and IFNgamma-R antigen expression. It also inhibited cleaved caspase 8 and 9 expression, decreased E-cadherin, and increased BclxL and cyclooxygenase-2 expression. The cDNA microarray approach showed that TGFbeta up-regulated the expression of genes with defined roles in tumoral progression sometimes associated with poor outcome in bladder cancer. CONCLUSION: These results suggest that a part of the bladder tumoral progression process may be related to the action of exogenous TGFbeta confirming the possible role for the microenvironment.
Subject(s)
Transforming Growth Factor beta/metabolism , Urinary Bladder Neoplasms/metabolism , Apoptosis/physiology , Cadherins/genetics , Cadherins/metabolism , Caspase 8 , Caspase 9 , Caspases/metabolism , Cell Differentiation , Cell Line, Tumor , Cell Proliferation , Disease Progression , Gene Expression Regulation, Neoplastic , Humans , Interferon-gamma/metabolism , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Protein Serine-Threonine Kinases , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-akt/metabolism , Receptor, Transforming Growth Factor-beta Type II , Receptors, Interferon/genetics , Receptors, Interferon/metabolism , Receptors, Transforming Growth Factor beta/genetics , Receptors, Transforming Growth Factor beta/metabolism , Receptors, Tumor Necrosis Factor, Type I/genetics , Receptors, Tumor Necrosis Factor, Type I/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Transforming Growth Factor beta/genetics , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/pathology , Interferon gamma ReceptorABSTRACT
BACKGROUND: TNFα and IFNγ, two main cytokines secreted in the urine of bladder cancer patients after Bacillus Calmette Guerin immunotherapy (BCG therapy), exert various responses ranging from growth arrest, apoptosis, phenotypic changes and differentiation. MATERIALS AND METHODS: To identify their transcriptional and translational targets, the highly sensitive bladder cancer cell line (RT112) was treated for 24 hours with increasing doses of IFNγ or TNFα and analyzed for cellular and molecular changes using a cDNA microarray technique (Transcriptome) containing 800 genes. RESULTS: High doses (>10 ng/ml) induced an apoptotic cell death, whereas low doses (<5 ng/ml) induced a survival program. TNFα-inducible genes, IFNγ-inducible genes and genes modulated by TNFα and IFNγ together were identified. All were related to the tumor progression program including cell proliferation, apoptosis/survival, angiogenesis and metastatic processes. CONCLUSION: These results suggest that the transcriptomic approach could be a good methodology to determine the molecular mechanisms involved in bladder tumor progression processes in relation to a low response to BCG treatment. However, mRNA and protein expression did not always correlate, suggesting that translational regulation is a vital process in bladder tumor progression.
