Open AccessProteomic changes of aryl hydrocarbon receptor (AhR)-silenced porcine granulosa cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)
Author(s) -
Karina Orlowska,
Sylwia Świgońska,
Agnieszka Sadowska,
Monika Ruszkowska,
Anynca,
Tomasz Molcan,
Agata Żmijewska,
Renata E. Ciereszko
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0223420
Subject(s) - aryl hydrocarbon receptor , chemistry , transfection , blot , protein disulfide isomerase , western blot , proteome , proteomics , receptor , intracellular , microbiology and biotechnology , annexin , biochemistry , biology , apoptosis , enzyme , transcription factor , gene
2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD) is a toxic man-made chemical compound contaminating the environment and affecting human/animal health and reproduction. Intracellular TCDD action usually involves the activation of aryl hydrocarbon receptor (AhR). The aim of the current study was to examine TCDD-induced changes in the proteome of AhR -silenced porcine granulosa cells. The AhR -silenced cells were treated with TCDD (100 nM) for 3, 12 or 24 h. Total protein was isolated, labeled with cyanines and next, the samples were separated by isoelectric focusing and SDS-PAGE. Proteins of interest were identified by MALDI-TOF/TOF mass spectrometry (MS) analysis and confirmed by western blotting and fluorescence immunocytochemistry. The AhR -targeted siRNA transfection reduced the granulosal expression level of AhR by 60–70%. In AhR -silenced porcine granulosa cells, TCDD influenced the abundance of only three proteins: annexin V, protein disulfide isomerase and ATP synthase subunit beta. The obtained results revealed the ability of TCDD to alter protein abundance in an AhR-independent manner. This study offers a new insight into the mechanism of TCDD action and provide directions for future functional studies focused on molecular effects exerted by TCDD.