Identification of toxicological biomarkers of di(2-ethylhexyl) phthalate in proteins secreted by HepG2 cells using proteomic analysis.

Seonyoung Choi, So Young Park, J. Jeong, Eunkyung Cho, Sohee Phark, Min Lee, Dongsub Kwak, Ji Youn Lim, Woon Won Jung, Dong Geun Sul

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The effects of di(2-ethylhexyl) phthalate (DEHP) on proteins secreted by HepG2 cells were studied using a proteomic approach. HepG2 cells were exposed to various concentrations of DEHP (0, 2.5, 5, 10, 25, 50, 100, and 250 microM) for 24 or 48 h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and comet assays were then conducted to determine the cytotoxicity and genotoxicity of DEHP, respectively. The MTT assay showed that 10 microM DEHP was the maximum concentration that did not cause cell death. In addition, the DNA damage in HepG2 cells exposed to DEHP was found to increase in a dose- and time-dependent fashion. Proteomic analysis using two different pI ranges (4-7 and 6-9) and large size 2-DE revealed the presence of 2776 protein spots. A total of 35 (19 up- and 16 down-regulated) proteins were identified as biomarkers of DEHP by ESI-MS/MS. Several differentiated protein groups were also found. Proteins involved in apoptosis, transportation, signaling, energy metabolism, and cell structure and motility were found to be up- or down-regulated. Among these, the identities of cystatin C, Rho GDP inhibitor, retinol binding protein 4, gelsolin, DEK protein, Raf kinase inhibitory protein, triose phosphate isomerase, cofilin-1, and haptoglobin-related protein were confirmed by Western blot assay. Therefore, these proteins could be used as potential biomarkers of DEHP and human disease associated with DEHP.

Original languageEnglish
Pages (from-to)1831-1846
Number of pages16
JournalProteomics
Volume10
Issue number9
DOIs
Publication statusPublished - 2010 May 1

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Hep G2 Cells
Biomarkers
Proteomics
Toxicology
Proteins
Assays
Cofilin 1
raf Kinases
Gelsolin
Triose-Phosphate Isomerase
Cystatin C
Retinol-Binding Proteins
Haptoglobins
Comet Assay
phthalic acid
Cell death
Protein Kinases
Cytotoxicity
Energy Metabolism
DNA Damage

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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Identification of toxicological biomarkers of di(2-ethylhexyl) phthalate in proteins secreted by HepG2 cells using proteomic analysis. / Choi, Seonyoung; Park, So Young; Jeong, J.; Cho, Eunkyung; Phark, Sohee; Lee, Min; Kwak, Dongsub; Lim, Ji Youn; Jung, Woon Won; Sul, Dong Geun.

In: Proteomics, Vol. 10, No. 9, 01.05.2010, p. 1831-1846.

Research output: Contribution to journalArticle

Choi, Seonyoung ; Park, So Young ; Jeong, J. ; Cho, Eunkyung ; Phark, Sohee ; Lee, Min ; Kwak, Dongsub ; Lim, Ji Youn ; Jung, Woon Won ; Sul, Dong Geun. / Identification of toxicological biomarkers of di(2-ethylhexyl) phthalate in proteins secreted by HepG2 cells using proteomic analysis. In: Proteomics. 2010 ; Vol. 10, No. 9. pp. 1831-1846.
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AU - Phark, Sohee

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