Toxicological biomarkers of 2,3,4,7,8-pentachlorodibenzofuran in proteins secreted by HepG2 cells

Sohee Phark, So Young Park, Seonyoung Choi, Zhi Zheng, Eunkyung Cho, Min Lee, Ji Youn Lim, Jong Bok Seo, Nam Hee Won, Woon Won Jung, Dong Geun Sul

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Abstract

Using a proteomic approach, a study was conducted for determination of the effects of 2,3,4,7,8-pentachlorodibenzofuran (2,3,4,7,8-PCDF) on proteins secreted by HepG2 cells. Briefly, HepG2 cells were exposed to various concentrations of 2,3,4,7,8-PCDF for 24 or 48 h. MTT and comet assays were then conducted for determination of cytotoxicity and genotoxicity, respectively. Results of an MTT assay showed that 1 nM of 2,3,4,7,8-PCDF was the maximum concentration that did not cause cell death. In addition, a dose- and time dependent increase of DNA damage was observed in HepG2 cells exposed to 2,3,4,7,8-PCDF. Therefore, two different concentrations of 2,3,4,7,8-PCDF, 1 and 5 nM, were selected for further analysis of proteomic biomarkers using two different pI ranges (4-7 and 6-9) and large two dimensional gel electrophoresis. Results showed identification of 32 proteins (29 up- and 3 down-regulated) by nano-LC-ESI-MS/MS and nano-ESI on a Q-TOF2 MS. Among these, the identities of pyridoxine-5'-phosphate oxidase, UDP-glucose 6-dehydrogenase, plasminogen activator inhibitor I precursor, plasminogen activator inhibitor-3, proteasome activator complex subunit 1, isoform 1 of 14-3-3 protein sigma, peptidyl-prolyl cis-trans isomerase A, 14-3-3 protein gamma, protein DJ-1, and nucleoside diphosphate kinase A were confirmed by western blot analysis. The differential expression of protein DJ-1, proteasome activator complex subunit 1 and plasminogen activator inhibitor-3 was further validated in plasma proteins from rats exposed to 2,3,4,7,8-PCDF. These proteins could be used as potential toxicological biomarkers of 2,3,4,7,8-PCDF.

Original languageEnglish
Pages (from-to)656-666
Number of pages11
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1824
Issue number4
DOIs
Publication statusPublished - 2012 Apr 1

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Keywords

  • 2,3,4,7,8-pentachlorodibenzofuran
  • Biomarker
  • HepG2 cell
  • Proteomic
  • Rat plasma
  • Two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Analytical Chemistry
  • Molecular Biology

Cite this

Phark, S., Park, S. Y., Choi, S., Zheng, Z., Cho, E., Lee, M., Lim, J. Y., Seo, J. B., Won, N. H., Jung, W. W., & Sul, D. G. (2012). Toxicological biomarkers of 2,3,4,7,8-pentachlorodibenzofuran in proteins secreted by HepG2 cells. Biochimica et Biophysica Acta - Proteins and Proteomics, 1824(4), 656-666. https://doi.org/10.1016/j.bbapap.2012.01.013