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

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• 作者：Sohee Phark^a ; ¹ ; So-Young Park^b ; ¹ ; Seonyoung Choi^a ; Zhi Zheng^a ; Eunkyung Cho^a ; Min Lee^a ; Ji-youn Lim^a ; Jong Bok Seo^c ; Nam Hee Won^d ; Woon-Won Jung^e ; Donggeun Sul^a ; ^f ; ^{dsul@korea.ac.kr}
• 关键词：2 ; 3 ; 4 ; 7 ; 8-PCDF ; 2 ; 3 ; 4 ; 7 ; 8-pentachlorodibenzofuran ; MTT ; 3-(4 ; 5-Dimethyl thiazol-2-yl)-2 ; 5-diphenyltetrazolium bromide ; LC-ESI-MS/MS ; liquid chromatography electrospray ionization-tandem mass spectrometry ; TOF2 MS ; time-of-flight 2 mass spectrometry ; PCDFs
• 刊名：Biochimica et Biophysica Acta - Proteins and Proteomics
• 出版年：2012
• 期刊代码：161_15709639
• 类别：bio
• 出版时间：April, 2012
• 卷：1824
• 期：4
• 页码：656-666
• 文件大小：1162 K

摘要

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.