Abstract
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants of air, water and soil, and are produced by the incomplete combustion of organic materials. The International Agency for Research on Cancer has characterized PAHs as carcinogens. In this study, we investigated the effects of benzo(a)pyrene (B(a)P), which is the most carcinogenic member of the PAHs, on Jurkat cell protein by proteomic analysis. Jurkat cells were treated with various concentrations of B(a)P (0, 2.5, 5, 10, 20 or 40 μM) for 24 or 48 h and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium and lactate dehydrogenase assays were carried out to determine cytotoxicity and a Comet assay was used to determinate genotoxicity. The cytotoxicity assays showed that 2.5 μM of B(a)P was the maximal concentration that did not cause any toxicity, but nevertheless, at this level B(a)P produced significant DNA damage in Jurkat cells at 48 h. Proteomic analysis using three different p/ ranges and large two-dimensional gel electrophoresis showed 3427 protein spots. A total of 46 (13 up- and 33 down-regulated) proteins were identified as biomarkers of B(a)P and showed dose-dependent expressions in Jurkat T-cell line exposed to B(a)P. Of these, 27 protein spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Two functionally differentiated protein groups were found. The protein group involving apoptosis and tumor suppression were found to be up-regulated, and B(a)P down-regulated enzyme was involved in energy metabolism, DNA synthesis and in cell structure and motility.
| Original language | English |
|---|---|
| Pages (from-to) | 3514-3526 |
| Number of pages | 13 |
| Journal | Proteomics |
| Volume | 4 |
| Issue number | 11 |
| DOIs | |
| Publication status | Published - 2004 Nov 1 |
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Keywords
- Benzo(a)pyrene
- Jurkat cell
- Matrix-assisted laser desorption/ionization-time of flight mass spectrometry
- Two-dimensional gel electrophoresis
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Effects of benzo(a)pyrene on protein expression in Jurkat T-cells. / Oh, Sangnam; Im, Hosub; Oh, Eunha; Lee, Joohyun; Khim, Jin Young; Mun, Joohee; Kim, Yanghee; Lee, Eun Il; Kim, Joon; Sul, Dong Geun.
In: Proteomics, Vol. 4, No. 11, 01.11.2004, p. 3514-3526.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Effects of benzo(a)pyrene on protein expression in Jurkat T-cells
AU - Oh, Sangnam
AU - Im, Hosub
AU - Oh, Eunha
AU - Lee, Joohyun
AU - Khim, Jin Young
AU - Mun, Joohee
AU - Kim, Yanghee
AU - Lee, Eun Il
AU - Kim, Joon
AU - Sul, Dong Geun
PY - 2004/11/1
Y1 - 2004/11/1
N2 - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants of air, water and soil, and are produced by the incomplete combustion of organic materials. The International Agency for Research on Cancer has characterized PAHs as carcinogens. In this study, we investigated the effects of benzo(a)pyrene (B(a)P), which is the most carcinogenic member of the PAHs, on Jurkat cell protein by proteomic analysis. Jurkat cells were treated with various concentrations of B(a)P (0, 2.5, 5, 10, 20 or 40 μM) for 24 or 48 h and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium and lactate dehydrogenase assays were carried out to determine cytotoxicity and a Comet assay was used to determinate genotoxicity. The cytotoxicity assays showed that 2.5 μM of B(a)P was the maximal concentration that did not cause any toxicity, but nevertheless, at this level B(a)P produced significant DNA damage in Jurkat cells at 48 h. Proteomic analysis using three different p/ ranges and large two-dimensional gel electrophoresis showed 3427 protein spots. A total of 46 (13 up- and 33 down-regulated) proteins were identified as biomarkers of B(a)P and showed dose-dependent expressions in Jurkat T-cell line exposed to B(a)P. Of these, 27 protein spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Two functionally differentiated protein groups were found. The protein group involving apoptosis and tumor suppression were found to be up-regulated, and B(a)P down-regulated enzyme was involved in energy metabolism, DNA synthesis and in cell structure and motility.
AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants of air, water and soil, and are produced by the incomplete combustion of organic materials. The International Agency for Research on Cancer has characterized PAHs as carcinogens. In this study, we investigated the effects of benzo(a)pyrene (B(a)P), which is the most carcinogenic member of the PAHs, on Jurkat cell protein by proteomic analysis. Jurkat cells were treated with various concentrations of B(a)P (0, 2.5, 5, 10, 20 or 40 μM) for 24 or 48 h and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium and lactate dehydrogenase assays were carried out to determine cytotoxicity and a Comet assay was used to determinate genotoxicity. The cytotoxicity assays showed that 2.5 μM of B(a)P was the maximal concentration that did not cause any toxicity, but nevertheless, at this level B(a)P produced significant DNA damage in Jurkat cells at 48 h. Proteomic analysis using three different p/ ranges and large two-dimensional gel electrophoresis showed 3427 protein spots. A total of 46 (13 up- and 33 down-regulated) proteins were identified as biomarkers of B(a)P and showed dose-dependent expressions in Jurkat T-cell line exposed to B(a)P. Of these, 27 protein spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Two functionally differentiated protein groups were found. The protein group involving apoptosis and tumor suppression were found to be up-regulated, and B(a)P down-regulated enzyme was involved in energy metabolism, DNA synthesis and in cell structure and motility.
KW - Benzo(a)pyrene
KW - Jurkat cell
KW - Matrix-assisted laser desorption/ionization-time of flight mass spectrometry
KW - Two-dimensional gel electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=8744258677&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=8744258677&partnerID=8YFLogxK
U2 - 10.1002/pmic.200400981
DO - 10.1002/pmic.200400981
M3 - Article
C2 - 15529408
AN - SCOPUS:8744258677
VL - 4
SP - 3514
EP - 3526
JO - Proteomics
JF - Proteomics
SN - 1615-9853
IS - 11
ER -