Toxicological biomarkers of 2,3,4,7,8-pentachlorodibenzofuran in proteins secreted by HepG2 cells.
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 48h. MTT and comet assays were then conducted for determination of cytotoxicity and genotoxicity, respectively. Results of an MTT assay showed that 1nM 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 5nM, 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.
Graduate School of Medicine, Korea University, Seoul, Republic of Korea.
Sohee
S
Phark
So-Young
SY
Park
Seonyoung
S
Choi
Zhi
Z
Zheng
Eunkyung
E
Cho
Min
M
Lee
Ji-youn
JY
Lim
Jong Bok
JB
Seo
Nam Hee
NH
Won
Woon-Won
WW
Jung
Donggeun
D
Sul
D000818:Animals; D001572:Benzofurans / Q000633:toxicity*; D015415:Biomarkers / Q000097:blood / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D004249:DNA Damage; D015180:Electrophoresis, Gel, Two-Dimensional; D004785:Environmental Pollutants / Q000633:toxicity*; D056945:Hep G2 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000097:blood / Q000378:metabolism; D008297:Male; D015513:Oncogene Proteins / Q000097:blood / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000097:blood / Q000378:metabolism; D018046:Protein C Inhibitor / Q000097:blood / Q000378:metabolism; D000071617:Protein Deglycase DJ-1; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization; D015854:Up-Regulation / Q000187:drug effects
D001572:Benzofurans; D015415:Biomarkers; D004785:Environmental Pollutants; D047908:Intracellular Signaling Peptides and Proteins; D015513:Oncogene Proteins; D018046:Protein C Inhibitor; D020543:Proteome; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D046988:Proteasome Endopeptidase Complex; C038890:2,3,4,7,8-pentachlorodibenzofuran