hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics

Wieser, Matthias; Stadler, Guido; Jennings, Paul; Streubel, Berthold; Pfaller, Walter; Ambros, Peter F.; Riedl, Claus R.; Katinger, Hermann; Grillari, Johannes; Grillari‐Voglauer, Regina

doi:10.1152/ajprenal.90405.2008

Cited by 269 publications

(251 citation statements)

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“…[22][23][24] Isolation and identification of rat primary tubular epithelial cells were performed according to previously published techniques, and cytokeratin 18 was used as a specific marker of tubular epithelial cells. [25][26][27] Interstitial fibroblasts were isolated and identified as reported. 28 FSP-1 was used as marker for renal fibroblasts.…”

Section: Isolation and Identification Of Primary Renal Residential Cellsmentioning

confidence: 99%

miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

Bai

Ding

et al. 2011

Journal of the American Society of Nephrology

193

145

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The molecular basis for aging of the kidney is not well understood. MicroRNAs (miRNAs) contribute to processes such as development, differentiation, and apoptosis, but their contribution to the aging process is unknown. Here, we analyzed the miRNA expression profile of young (3-month) and old (24-month) rat kidneys and identified the biologic pathways and genes regulated by differentially expressed miRNAs. We observed upregulation of 18 miRNAs with aging, mainly regulating the genes associated with energy metabolism, cell proliferation, antioxidative defense, and extracellular matrix degradation; in contrast, we observed downregulation of 7 miRNAs with aging, principally targeting the genes associated with the immune inflammatory response and cell-cycle arrest. Bioinformatics analysis suggested that superoxide dismutase 2 (SOD2) and thioredoxin reductase 2 (Txnrd2), located in the mitochondria, are potential targets of miR-335 and miR-34a, respectively. Aging mesangial cells exhibited significant upregulation of miR-335 and miR-34a and marked downregulation of SOD2 and Txnrd2. miR-335 and miR-34a inhibited expression of SOD2 and Txnrd2 by binding to the 3Ј-untranslated regions of each gene, respectively. Overexpression of miR-335 and miR-34a induced premature senescence of young mesangial cells via suppression of SOD2 and Txnrd2 with a concomitant increase in reactive oxygen species (ROS). Conversely, antisense miR-335 and miR-34a inhibited senescence of old mesangial cells via upregulation of SOD2 and Txnrd2 with a concomitant decrease in ROS. In conclusion, these results suggest that miRNAs may contribute to renal aging by inhibiting intracellular pathways such as those involving the mitochondrial antioxidative enzymes SOD2 and Txnrd2. 22: 125222: -126122: , 201122: . doi: 10.1681 Kidney aging is an important clinical problem, not only because normal aging reduces renal function but also because of the high frequency of ESRD, renal cancer, and renal failure in elderly people. Renal aging is of interest as a general model for organ aging because renal function can be quantitatively assessed more readily than that of other organs in clinical practice. 1 At the present time, the molecular basis of renal aging is not clearly known. For example, nothing is known of the role of microRNAs (miRNAs) in the aging process of organs. J Am Soc NephrolmiRNAs are a novel class of small, regulatory, noncoding RNA molecules that inhibit the expression of multiple genes at the post-transcriptional level. miRNAs have been found to play a crucial role in development, differentiation, apoptosis, and metabolism and are involved in the pathogenesis of many human diseases. 2,3 Bioinformatics studies suggest that miRNAs may regulate Ͼ60% of all human genes. 4,5 Studies have shown that overexpression of miRNA lin-4 increases longevity in Caenorhabditis elegans, whereas loss of lin-4 leads to a

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Section: Isolation and Identification Of Primary Renal Residential Cellsmentioning

confidence: 99%

miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

Bai

Ding

et al. 2011

Journal of the American Society of Nephrology

193

145

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“…Normal human renal proximal tubular epithelial cells (nRPteCs) and human exfoliated proximal tubular epithelial cells (HePteCs) were isolated from urine samples as described in detail previously (Wieser et al, 2008(Wieser et al, , 2014 and cultivated in ProxUp-Pri medium (evercyte GmbH).…”

Section: Commercial Cell Linesmentioning

confidence: 99%

“…even though very valuable, these models are hampered due to limited population doublings, resulting in the need for large amounts of human tissue samples from which the cells must be prepared and therefore high variability in RPteC cell behavior between isolations from different donors. Also, the human kidney cell line HK-2, immortalized by a recombinant retrovirus containing the human papilloma virus e6/e7 genes, has been used, but controversial results concerning this cell line's RPteC-like characteristics, including drug transporter expression, have been published (Ryan et al, 1994;Wieser et al, 2008;Jenkinson et al, 2012). Moreover, for all cell lines used for toxicity testing so far alterations of the primary cell-like characteristics due to viral oncogenes used for immortalization were noted and are seen as major shortfalls (Stacey and Hartung, 2006), even though the use of conditional SV40 in combination with telomerase seems promising (Wilmer et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…Still, telomerase alone is regarded as the most appropriate strategy to immortalize cells without altering the primary cell-like key functionalities, and a large variety of different human cells have been successfully immortalized using hteRt (Bodnar et al, 1998;Chang et al, 2005). We have previously reported that introduction of hteRt alone is sufficient for immortalization and does not change the functional characteristics of epithelial cells of renal proximal tubules (Wieser et al, 2008). However, it may prove useful to employ a panel of cell lines originating from more than one individual to represent different genetic backgrounds and so improve predictivity.…”

Section: Introductionmentioning

confidence: 99%

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Controversial role of Gamma-Glutamyl Transferase activity in cisplatin nephrotoxicity

Fliedl

2014

ALTEX

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“…In addition to rat cells, we used human proximal tubule cells RPTEC/TERT1 16 as currently it is not clear if MON is a rat-specific carcinogen.…”

Section: Introductionmentioning

confidence: 99%

Transcriptomic alterations induced by Monuron in rat and human renal proximal tubule cells in vitro and comparison to rat renal-cortex in vivo

et al. 2015

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-dimethyl-3-(4-chlorophenyl)urea) is a non-selective phenylurea herbicide, widely used in developing countries although concerns have been raised about its toxicity and carcinogenicity. Monuron was evaluated by the National Toxicology Program in 1988 and shown to be a male rat-specific renal carcinogen. We report that oral administration of Monuron to male rats for 3 days, leads to a larger number of genes being differentially expressed in the renal-cortex than in the liver. Further, we observed up-regulation of cell cycle genes and genes involved in cell proliferation in the renalcortex while in the liver xenobiotic metabolising enzymes were up-regulated. We also identified one commonly down-regulated gene in both organs -fragile histidine triad gene (Fhit), a putative tumour suppressor gene; however the down-regulation was only significant at the protein level in the liver. In addition, we conducted in vitro wholegenome transcriptomics studies with human and rat renal cortical cells. Rat cells exposed to Monuron showed down-regulation of sterol biosynthesis, spliceosome and cell cycle genes and up-regulation of genes involved in amino acid metabolism and transport. No genes were found to be differentially expressed in human cells exposed to Monuron. Overall, the findings from the in vitro studies showed very little overlap with the whole animal findings.

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hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics

Cited by 269 publications

References 68 publications

miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

Controversial role of Gamma-Glutamyl Transferase activity in cisplatin nephrotoxicity

Transcriptomic alterations induced by Monuron in rat and human renal proximal tubule cells in vitro and comparison to rat renal-cortex in vivo

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