H-ras-transformed NRK-52E renal epithelial cells have altered growth, morphology, and cytoskeletal structure that correlates with renal cell carcinoma in vivo

Best, Carolyn J.M.; Tanzer, Lee R.; Phelps, Patricia C.; Merriman, Ronald L.; Boder, George G.; Trump, Benjamin F.; Elliget, Kathryn A.

doi:10.1007/s11626-999-0028-2

Cited by 16 publications

(12 citation statements)

References 48 publications

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“…RCCs are extremely resistant to conventional cancer therapies, and no effective systemic therapy presently exists for patients with metastatic RCCs. Although some apoptosis-related genes have altered expression in RCCs, this appears to relate largely to genes that have functions in the broader context of apoptosis induction, such as H-Ras [29], rather than genes whose products make up the apoptosis pathway itself, for example, the Bcl-2-gene family. Kolenko et al [22]have demonstrated in some human-derived RCC cell lines that caspase-3 expression is low or lacking.…”

Section: Discussionmentioning

confidence: 99%

Comparative Analysis of Caspase Activation and Apoptosis in Renal Tubular Epithelial Cells and Renal Cell Carcinomas

Davidson

Percy

Rennick

et al. 2005

Nephron Exp Nephrol

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Background/Aims: Treatment of renal cell carcinoma (RCC) is limited by its resistance to conventional chemotherapies. This may occur, in part, from resistance to apoptosis. The role of caspase activation in apoptosis resistance in treated RCCs was investigated. Methods: Two human RCC cell lines (ACHN and SN12K1) and renal tubular epithelial cells (HK2) were treated with 5-fluorouracil (0.2–20 µg/ml) or cisplatin (1–100 µM). Activation of caspase-3 and -2 was analysed and compared with levels of apoptosis. Caspase function was analysed using pan-caspase inhibition (z-VAD-fmk) and caspase-2 inhibition (z-VDVAD-fmk). Results: RCC apoptosis was significantly lower (p < 0.05) than in HK2s after treatment, confirming their chemoresistance. Pro-caspase-3 (32 kDa) was detected in all cell lines. Cleaved caspase-3 (19 kDa) was not detected by Western immunoblots in treated RCCs and only minimal activated caspase-3 was detected in treated RCCs using immunohistochemistry. All cells had pro-caspase-2 (48 kDa) and the activated form (33 kDa) appeared in all treated cells. Caspase inhibition caused a reduction in, but not negation of, therapy-induced apoptosis in HK2s and RCCs (p < 0.05 for HK2s and ACHN cells), indicating that a caspase activation pathway must occur in RCC apoptosis but this pathway does not act via caspase-3 cleavage. Inhibition of caspase-2 reduced apoptosis only in HK2s, indicating that the activated caspase-2, identified in treated RCCs, was not responsible for their apoptosis induction. Conclusion: Specific differences in caspase-3 and -2 activation were identified in renal tubular epithelium and RCCs after chemotherapy. Identification of RCC-specific caspase inactivation or redundancy may explain, in part, the resistance of RCCs to cancer therapies and may be useful in targeting apoptotic pathways to overcome RCC resistance to treatment.

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Section: Discussionmentioning

confidence: 99%

Comparative Analysis of Caspase Activation and Apoptosis in Renal Tubular Epithelial Cells and Renal Cell Carcinomas

Davidson

Percy

Rennick

et al. 2005

Nephron Exp Nephrol

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“…This cell line, derived from the proximal tubule epithelium (PTE) of the rat kidney, is non-tumorigenic, and retains many morphological and physiological characteristics of PTE in vivo [18, 19]. Cells were incubated in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum and antibiotics (100 U/ml penicillin, 100 µg/ml streptomycin) at 37°C in 95% O 2 /5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Inhibition of c-Jun N-Terminal Kinase Ameliorates Apoptosis Induced by Hydrogen Peroxide in the Kidney Tubule Epithelial Cells (NRK-52E)

Wang¹,

Matsushita²,

Araki³

et al. 2002

Nephron

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Background: Hydrogen peroxide (H₂O₂)-induced apoptosis has been shown to be involved in ischemic and toxic tubular damage. Recent studies have revealed that oxidative stress can activate c-Jun N-terminal kinase (JNK), and the oxidative stress-JNK pathway is an important apoptotic pathway of cells exposed to various stresses. The present study was designed to investigate JNK activation and the effects of the JNK pathway inhibition during H₂O₂-induced apoptosis in kidney epithelial tubule cells (NRK-52E). Materials and Methods: NRK-52E cells were treated with 0–500 µM H₂O₂ and/or 100 µM quercetin (an inhibitor of the JNK pathway). Apoptosis was assessed by flow cytometry analysis and DNA ladder. JNK activity was assessed by the GST-c-Jun (1-169) binding/protein kinase assay. Results: H₂O₂ induced apoptosis in NRK-52E cells in a concentration-dependent manner, which was demonstrated by the reduced DNA PI staining, externalization of phosphatidylserine and DNA ladder. Apoptosis induced by H₂O₂was accompanied by JNK activation and up-regulated JNK activity. Quercetin treatment suppressed the JNK activity and ameliorated apoptosis induced by H₂O₂. Conclusions: H₂O₂ induced apoptosis in NRK-52E cells, which was associated with activation and up-regulation of JNK. Quercetin treatment could decrease JNK activity and ameliorate H₂O₂-induced apoptosis.

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“…TSP-1 at 1 µg/ml was added to cultured kidney proximal tubule (NRK) cells and studied at 24 and 48 hours for evidence of apoptosis by cytochrome c release and caspase-3 activity. The NRK-52E normal kidney cell line is derived from the proximal tubule epithelium (PTE) of the rat kidney, is nontumorigenic, retains many morphological and physiological characteristics of PTE, and has been used to mimic changes in vivo (31)(32)(33). Cytochrome c apoptosis detection kit (Alexa Fluor 488 assay; Invitrogen Corp.) was used to perform the apoptosis assay.…”

Section: Kidney Ir Injury: Animal Model Kidney Ir Injury Was Inducedmentioning

confidence: 99%

Identification of thrombospondin 1 (TSP-1) as a novel mediator of cell injury in kidney ischemia

Thakar

Zahedi

Revelo

et al. 2005

Journal of Clinical Investigation

122

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Thrombospondin 1 (TSP-1) is a matricellular protein that inhibits angiogenesis and causes apoptosis in vivo and in vitro in several cancerous cells and tissues. Here we identify TSP-1 as the molecule with the highest induction level at 3 hours of IR injury in rat and mouse kidneys subjected to ischemia/reperfusion (IR) injury using the DNA microarray approach. Northern hybridizations demonstrated that TSP-1 expression was undetectable at baseline, induced at 3 and 12 hours, and returned to baseline levels at 48 hours of reperfusion. Immunocytochemical staining identified the injured proximal tubules as the predominant sites of expression of TSP-1 in IR injury and showed colocalization of TSP-1 with activated caspase-3. Addition of purified TSP-1 to normal kidney proximal tubule cells or cells subjected to ATP depletion in vitro induced injury as demonstrated by cytochrome c immunocytochemical staining and caspase-3 activity. The deleterious role of TSP-1 in ischemic kidney injury was demonstrated directly in TSP-1 null mice, which showed significant protection against IR injury-induced renal failure and tubular damage. We propose that TSP-1 is a novel regulator of ischemic damage in the kidney and may play an important role in the pathophysiology of ischemic kidney failure.

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H-ras-transformed NRK-52E renal epithelial cells have altered growth, morphology, and cytoskeletal structure that correlates with renal cell carcinoma in vivo

Cited by 16 publications

References 48 publications

Comparative Analysis of Caspase Activation and Apoptosis in Renal Tubular Epithelial Cells and Renal Cell Carcinomas

Comparative Analysis of Caspase Activation and Apoptosis in Renal Tubular Epithelial Cells and Renal Cell Carcinomas

Inhibition of c-Jun N-Terminal Kinase Ameliorates Apoptosis Induced by Hydrogen Peroxide in the Kidney Tubule Epithelial Cells (NRK-52E)

Identification of thrombospondin 1 (TSP-1) as a novel mediator of cell injury in kidney ischemia

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