Proteomic analysis of proteins secreted by HepG2 cells treated with butyl benzyl phthalate.
Proteomic changes in proteins secreted by human hepatocellular carcinomas (HepG2) cells exposed to butyl benzyl phthalate (BBP) were evaluated. HepG2 cells were treated with three different concentrations of BBP (0, 10, or 25 μM) for 24 or 48 h. Following incubation, the cells were subjected to proteomic analysis using two different pI ranges (4-7 and 6-9) and large-size two-dimensional gel electrophoresis. Results showed resolution of a total of 2776 protein spots. Of these, 29, including 19 upregulated and 10 downregulated proteins, were identified by electrospray ionization-mass spectrometry-mass spectrometry (ESI-MS/MS). Among these, the identities of cystatin C, Rho guanine nucleotide dissociation inhibitor, gelsolin, DEK protein, Raf kinase inhibitory protein, triose phosphate isomerase, heptaglobin-related protein, inter-alpha-trypsin inhibitor heavy chain H2, and electron transfer flavoprotein subunit beta were confirmed by Western blot analysis. These proteins were found to be involved in apoptosis, signaling, tumor progression, energy metabolism, and cell structure and motility. Therefore, these proteins have potential to be employed as biomarkers of BBP exposure and may be useful in understanding mechanisms underlying the adverse effects of BBP.
Choi
Seonyoung
S
Department of Nanobiomedical Science, College of Advanced Science, Chung-Nam, Korea.
Park
So-Young
SY
Kwak
Dongsub
D
Phark
Sohee
S
Lee
Min
M
Lim
Ji-Youn
JY
Jung
Woon-Won
WW
Sul
Donggeun
D
D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D004249:DNA Damage; D015180:Electrophoresis, Gel, Two-Dimensional; D005562:Formazans / Q000378:metabolism; D056945:Hep G2 Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D008113:Liver Neoplasms / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D046228:Microarray Analysis; D010449:Peptide Mapping; D010795:Phthalic Acids / Q000493:pharmacokinetics / Q000633:toxicity*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley; D016622:Silver Staining; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D013723:Teratogens / Q000493:pharmacokinetics / Q000633:toxicity*; D013778:Tetrazolium Salts / Q000378:metabolism
D014408:Biomarkers, Tumor; D005562:Formazans; D010795:Phthalic Acids; D020543:Proteome; D013723:Teratogens; D013778:Tetrazolium Salts; C079782:MTT formazan; C027561:butylbenzyl phthalate