Archives of Toxicology

, Volume 89, Issue 11, pp 1971–1979 | Cite as

Increased susceptibility of H-Ras G12V -transformed human urothelial cells to the genotoxic effects of sodium arsenite

Inorganic Compounds
First Online:
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Accepted:

Abstract

Inorganic arsenite (iAs) is a human carcinogen. Numerous studies have shown that mutation-activated H-Ras is frequently observed in human urothelial carcinomas. The interaction between iAs, an environmental factor, and H-Ras, an oncogene, is not clear. In this study, we explored the genotoxic effects of iAs in human urothelial cells ectopically expressing H-Ras G12V , an activated H-Ras oncogene. Our results showed that H-Ras G12V -transformed human urothelial cells (HUC-RAS) were more susceptible to arsenite-induced cell death, DNA damage, micronuclei formation and anchorage-independent growth than control cells (HUC-neo). Furthermore, iAs treatment induced higher intracellular levels of reactive oxygen species (ROS) in the HUC-RAS cells than in the HUC-neo cells. N-acetyl-l-cysteine could suppress the iAs-induced increases in ROS and genetic damage. We further demonstrated that the intracellular glutathione levels were significantly elevated by the iAs treatment of the HUC-neo cells, but that this effect was not observed in the HUC-RAS cells. The iAs treatment induced higher superoxide dismutase activity in the HUC-neo cells than in the HUC-RAS cells. Alternatively, catalase activity was higher in the HUC-RAS cells than in the HUC-neo cells, but this enzyme was significantly suppressed by iAs. Moreover, iAs activated the ERK and JNK signaling pathways, which are involved in iAs-induced ROS production and genetic damage. Taken together, our present results suggest that elevated catalase activity in H-Ras G12V -transformed cells is significantly suppressed by iAs via activation of ERK and JNK signaling pathways and hence attenuate the defense of the neoplastic transformed cells against iAs-induced oxidative injuries.

Keywords

Arsenic H-Ras oncogene Genotoxicity Reactive oxygen species Catalase Stress signaling 

Abbreviations

iAs

Inorganic arsenite

MMA

Monomethylarsonic acid

DMA

Dimethylarsinic acid

ROS

Reactive oxygen species

MAPK

Mitogen-activated protein kinase

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Notes

Acknowledgments

We are grateful to Dr. Pin-pin Lin (Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Zhunan, Taiwan, ROC) for technical support in the ICP-MS analysis of arsenic contents and experimental assistance. We also thank Dr. Y. F. Liao (Institute of Cellular and Organism Biology, Academia Sinica, Taipei, Taiwan, ROC) for providing us with the pCMV vector. The authors thank the Pathological Core Laboratory, which is supported by the Institute of Biomedical Sciences at the Academia Sinica. This work was supported by the Academia Sinica and Grants from the National Science Council, Taiwan, ROC (NSC97-2314-B-001-003-MY3 and NSC98–2320-B-001–002-MY3).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

204_2014_1344_MOESM1_ESM.pdf (10 kb)
Supplementary material 1 (PDF 11 kb)
204_2014_1344_MOESM2_ESM.pdf (431 kb)
Supplementary material 2 (PDF 432 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Institute of PharmacologyNational Yang-Ming UniversityTaipeiTaiwan
  2. 2.Institute of Biomedical SciencesAcademia SinicaTaipeiTaiwan

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