Aristolochic acid suppresses DNA repair and triggers oxidative DNA damage in human kidney proximal tubular cells

Ya Yin Chen, Jing-Gung Chung, Hsiu Ching Wu, Da-Tian Bau, Kuen Yuh Wu, Shung Te Kao, Chien Yun Hsiang, Tin Yun Ho, Su Yin Chiang

Research output: Contribution to journalArticle

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Abstract

Aristolochic acid (AA), derived from plants of the Aristolochia genus, has been proven to be associated with aristolochic acid nephropathy (AAN) and urothelial cancer in AAN patients. In this study, we used toxicogenomic analysis to clarify the molecular mechanism of AA-induced cytotoxicity in normal human kidney proximal tubular (HK-2) cells, the target cells of AA. AA induced cytotoxic effects in a dose-dependent (10, 30, 90 μM for 24 h) and time-dependent manner (30 μM for 1, 3, 6, 12 and 24 h). The cells from those experiments were then used for microarray experiments in triplicate. Among the differentially expressed genes analyzed by Limma and Ingenuity Pathway Analysis software, we found that genes in DNA repair processes were the most significantly regulated by all AA treatments. Furthermore, response to DNA damage stimulus, apoptosis, and regulation of cell cycle, were also significantly regulated by AA treatment. Among the differentially expressed genes found in the dose-response and time-course studies that were involved in these biological processes, two up-regulated (GADD45B, NAIP), and six down-regulated genes (TP53, PARP1, OGG1, ERCC1, ERCC2, and MGMT) were confirmed by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). AA exposure also caused a down-regulation of the gene expression of antioxidant enzymes, such as superoxide dismutase, glutathione reductase, and glutathione peroxidase. Moreover, AA treatment led to increased frequency of DNA strand breaks, 8-hydroxydeoxyguanosine-positive nuclei, and micronuclei in a dose-dependent manner in HK-2 cells, possibly as a result of the inhibition of DNA repair. These data suggest that oxidative stress plays a role in the cytotoxicity of AA. In addition, our results provide insight into the involvement of down-regulation of DNA repair gene expression as a possible mechanism for AA-induced genotoxicity.

Original languageEnglish (US)
Pages (from-to)141-153
Number of pages13
JournalOncology reports
Volume24
Issue number1
DOIs
StatePublished - Jan 1 2010

Fingerprint

DNA Repair
DNA Damage
Kidney
aristolochic acid I
Down-Regulation
Aristolochia
Toxicogenetics
Genes
Biological Phenomena
DNA Breaks
Glutathione Reductase
p53 Genes
Gene Expression Regulation
Glutathione Peroxidase
Reverse Transcription
Reaction Time
Superoxide Dismutase
Cell Cycle
Oxidative Stress
Therapeutics

Keywords

  • Aristolochic acid
  • DNA repair
  • Oxidative DNA damage

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Aristolochic acid suppresses DNA repair and triggers oxidative DNA damage in human kidney proximal tubular cells. / Chen, Ya Yin; Chung, Jing-Gung; Wu, Hsiu Ching; Bau, Da-Tian; Wu, Kuen Yuh; Kao, Shung Te; Hsiang, Chien Yun; Ho, Tin Yun; Chiang, Su Yin.

In: Oncology reports, Vol. 24, No. 1, 01.01.2010, p. 141-153.

Research output: Contribution to journalArticle

Chen, Ya Yin ; Chung, Jing-Gung ; Wu, Hsiu Ching ; Bau, Da-Tian ; Wu, Kuen Yuh ; Kao, Shung Te ; Hsiang, Chien Yun ; Ho, Tin Yun ; Chiang, Su Yin. / Aristolochic acid suppresses DNA repair and triggers oxidative DNA damage in human kidney proximal tubular cells. In: Oncology reports. 2010 ; Vol. 24, No. 1. pp. 141-153.
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