Effects of benzo(a)pyrene on protein expression in Jurkat T‐cells

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 microM) 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 microM 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 pI 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.

Evaluation of plasma carcinogenic markers in rat hepatic tumors models induced by rat hepatoma N1-S1 cells and benzo[a]pyrene

“…Plasma samples were prepared for 2-DE analysis as described previously (Oh et al, 2004). Lipids and salts were removed from samples using a molecular weight cut off column (3 kDa; Amicon, Millipore).…”

“…This reactive metabolite exerts mutagenic and carcinogenic effects by reacting with DNA to form miscoding adducts, leading to the initiation of carcinogenesis (Conklin, 2000;Gelboin, 1980;Hicks, 1983). Previously, we reported that BaP down-regulates the expression of breast cancer metastasis-suppressor 1, differentiation-related gene (Drg1, a putative tumor suppressor) and a putative lung tumor suppressor in Jurkat cells (Oh et al, 2004). Furthermore, the increased DNA damage induced by BaP has been associated with the activation of p53 tumor suppressor and induction of programmed cell death (Umannova et al, 2008).…”

Binary Mixtures of Polycyclic Aromatic Hydrocarbons Display Nonadditive Mixture Interactions in an In Vitro Liver Cell Model

“…Following BaP exposure, some cells significantly downregulate proteins that are involved in metastasis and tumor suppression . For example, one study found that downregulated proteins were involved with apoptosis, cell structure, metabolism, and DNA synthesis . Proteins involved with cell proliferation, growth, and differentiation were also upregulated .…”

Contributions to our understanding of T cell physiology through unveiling the T cell proteome

“…A decrease in proteins associated with DNA synthesis was also found by Oh et al . [44] investigating benzo(a)pyrene [B(a)P], a polycyclic aromatic hydrocarbon which is a ubiquitous pollutant and carcinogen. Two concentrations of B(a)P, one cytotoxic, were used to investigate the potential for detection of biomarkers of diseases associated with B(a)P. Proteins associated with apoptosis and tumour suppression were found to be up‐regulated and those associated with energy metabolism, DNA synthesis and cell structure were down‐regulated.…”