Immortalisation of human urothelial cells

Petzoldt, J. L.; Leigh, I. M.; Duffy, P.G.; Sexton, Connie J.; Masters, J. R. W.

doi:10.1007/bf00698738

Cited by 90 publications

(87 citation statements)

References 7 publications

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“…Mary Ann and Donald Sens (University of North Dakota). UROtsa cells were grown in DMEM medium enriched with 5% FBS according to the original condition described (Petzoldt et al, 1995). All mammalian cells were incubated at 37° C in a humidified incubator containing 5% CO 2 .…”

Section: Chemicals Cell Culture Transfection and Inductionmentioning

confidence: 99%

Activation of Nrf2 by arsenite and monomethylarsonous acid is independent of Keap1-C151: enhanced Keap1–Cul3 interaction

Wang

Sun

Chen

et al. 2008

Toxicology and Applied Pharmacology

126

101

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Drinking water contaminated with arsenic, a human carcinogen, is a worldwide health issue. An understanding of cellular signaling events in response to arsenic exposure and rational designing of strategies to reduce arsenic damages by modulating signaling events are important to fight against arsenic-induced diseases. Previously, we reported that activation of the Nrf2-mediated cellular defense pathway confers protection against toxic effects induced by sodium arsenite [As(III)] or monomethylarsonous acid [MMA(III)]. Paradoxically, arsenic has been reported to induce the Nrf2-dependent signaling pathway. Here, we report the unique mechanism of Nrf2 induction by arsenic. Similar to tert-butylhydroquinone (tBHQ) or sulforaphane (SF), arsenic induced the Nrf2-dependent response through enhancing Nrf2 protein levels by inhibiting Nrf2 ubiquitination and degradation. However, the detailed action of arsenic in Nrf2 induction is different from that of tBHQ or SF. Arsenic markedly enhanced the interaction between Keap1 and Cul3, subunits of the E3 ubiquitin ligase for Nrf2, which led to impaired dynamic assembly/disassembly of the E3 ubiquitin ligase and thus decreased its ligase activity. Furthermore, induction of Nrf2 by arsenic is independent of the previously identified C151 residue in Keap1 that is required for Nrf2 activation by tBHQ or SF. Distinct mechanisms of Nrf2 activation by seemingly harmful and beneficial reagents provide a molecular basis to design Nrf2-activating agents for therapeutic intervention.

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Section: Chemicals Cell Culture Transfection and Inductionmentioning

confidence: 99%

Activation of Nrf2 by arsenite and monomethylarsonous acid is independent of Keap1-C151: enhanced Keap1–Cul3 interaction

Wang

Sun

Chen

et al. 2008

Toxicology and Applied Pharmacology

126

101

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“…46 J82 is derived from an invasive, poorly differentiated high-grade urothelial 8p deletions and sFRP1 loss in bladder cancer R Stoehr et al carcinoma of the bladder. 47 For comparison, the cell line UROtsa, derived from normal urothelium by immortalization with SV40 large T antigen, 48 was investigated. Cells were maintained in RPMI supplemented with 1% L-glutamine, 1% sodium pyruvate and 5% fetal calf serum at 371C in a humidified atmosphere containing 5% CO 2 .…”

Section: Cell Linesmentioning

confidence: 99%

Deletions of chromosome 8p and loss of sFRP1 expression are progression markers of papillary bladder cancer

Stoehr

Wissmann²,

Suzuki³

et al. 2004

Laboratory Investigation

134

101

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Many molecular alterations are known to occur in urothelial carcinoma of the bladder, but their significance for tumor progression is poorly understood. Deletions of chromosome 8p are frequently found in several tumor types and are often associated with progressive disease. In all, 99 bladder tumors were screened for deletions at 8p using loss of heterozygosity (LOH) and multicolor fluorescence in situ hybridization FISH analyses. Allelic loss on chromosome 8p in at least one marker was found in 25/99 (25%) tumors. There was a significant correlation of 8p deletions with invasive tumor growth and a highly significant association with papillary growth pattern in patients with invasive disease. cDNA array analyses revealed that secreted Frizzled-related protein 1 (sFRP1), an antagonist of Frizzled receptors and Wnt pathway activation on chromosome 8p12-11.1, is frequently downregulated in bladder cancer. To investigate sFRP1 as a candidate for a putative progressionrelated gene on 8p, urothelial cell lines and primary urothelial carcinomas were screened for sFRP1 expression using quantitative real-time PCR, Northern blot, immunofluorescence and immunohistochemistry (IHC). Of the investigated bladder cancers, 38% showed loss of sFRP1 expression by quantitative RT-PCR. Evaluation of the protein expression by IHC using tissue microarrays containing 776 bladder cancers revealed loss or strong reduction of sFRP1 expression in 66% of cases. SFRP1 loss was associated with higher tumor stage and grade and shorter overall survival. In addition, loss of sFRP1 was an independent indicator of poor survival in patients with papillary but not with muscle invasive bladder cancer. There were neither mutations in the coding region of sFRP1 nor homozygous deletions at 8p12-11.21. However, promoter methylation was detected using methylation-specific PCR in 29% of cases. In conclusion, we could show a close correlation of chromosome 8p deletions and progression of papillary bladder tumors. The sFRP1 gene on chromosome 8p12-11.1 could be a candidate gene for the predicted, progression-related tumor suppressor gene in bladder cancer and could contribute to urothelial carcinogenesis.

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“…The mRNA for the protein is expressed in a variety of rat tissues and in HepG2 cells, a human hepatoma cell line. However, it is not expressed in UROtsa cells, an SV40 large T antigen-immortalized human urothelial line (27), that do not methylate inorganic arsenic. (22).…”

mentioning

confidence: 99%

A Novel S-Adenosyl-l-methionine:Arsenic(III) Methyltransferase from Rat Liver Cytosol

Lin

Shi

Nix

et al. 2002

Journal of Biological Chemistry

318

216

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S-Adenosyl-L-methionine (AdoMet):arsenic(III) methyltransferase, purified from liver cytosol of adult male Fischer 344 rats, catalyzes transfer of a methyl group from AdoMet to trivalent arsenicals producing methylated and dimethylated arsenicals. The kinetics of production of methylated arsenicals in reaction mixtures containing enzyme, AdoMet, dithiothreitol, glutathione (GSH), and arsenite are consistent with a scheme in which monomethylated arsenical produced from arsenite is the substrate for a second methylation reaction that yields dimethylated arsenical. The mRNA for this protein predicts a 369-amino acid residue protein (molecular mass 41056) that contains common methyltransferase sequence motifs. Its sequence is similar to Cyt19, a putative methyltransferase, expressed in human and mouse tissues. Reverse transcription-polymerase chain reaction detects S-adenosyl-L-methionine:arsenic(III) methyltransferase mRNA in rat tissues and in HepG2 cells, a human cell line that methylates arsenite and methylarsonous acid. S-Adenosyl-L-methionine:arsenic-(III) methyltransferase mRNA is not detected in UROtsa cells, an immortalized human urothelial cell line that does not methylate arsenite. Because methylation of arsenic is a critical feature of its metabolism, characterization of this enzyme will improve our understanding of this metalloid's metabolism and its actions as a toxin and a carcinogen.In many species, including humans, exposure to inorganic arsenic results in urinary excretion of methylated and dimethylated arsenicals (1-3). Cullen and co-workers (4) summarized the conversion of inorganic arsenic into these methylated products in a reaction scheme which incorporates oxidative methylation and the cycling of arsenic between the pentavalent (As V ) 1 and trivalent (As III ) oxidation states,Because reduction of arsenic to trivalency is a prerequisite for its oxidative methylation, pentavalent arsenicals are reduced by endogenous thiols such as glutathione (GSH) (5, 6) or by As V reductases (7-9). A protein has been purified from rabbit liver cytosol that catalyzes the methylation of both arsenite and methylarsonous acid (10, 11); however, this protein has not been sequenced. These activities are designated arsenite methyltransferase (EC 2.1.1.137) and methylarsonite methyltransferase (EC 2.1.1.138), respectively. This protein (estimated molecular mass 60 kDa) uses S-adenosyl-L-methionine (AdoMet) as the methyl group donor. The methylation of arsenite by this protein is stimulated by a monothiol (GSH) and the methylation of methylarsonous acid is highly stimulated by a dithiol, dithiothreitol (DTT). The methylation of arsenic has been commonly regarded as a mechanism for its detoxification (12). However, recent research has shown that methylated arsenicals that contain As III are important intermediates in the metabolism of inorganic arsenic. Methylated arsenicals that contain As III are found in the urine of individuals who chronically consume drinking water that contains inorganic arsenic and in cells cu...

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Immortalisation of human urothelial cells

Cited by 90 publications

References 7 publications

Activation of Nrf2 by arsenite and monomethylarsonous acid is independent of Keap1-C151: enhanced Keap1–Cul3 interaction

Activation of Nrf2 by arsenite and monomethylarsonous acid is independent of Keap1-C151: enhanced Keap1–Cul3 interaction

Deletions of chromosome 8p and loss of sFRP1 expression are progression markers of papillary bladder cancer

A Novel S-Adenosyl-l-methionine:Arsenic(III) Methyltransferase from Rat Liver Cytosol

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