The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the proteome of porcine granulosa cells

Identification and characterization of long non-coding RNAs in porcine granulosa cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

“…NHS products, in turn, were described to maintain cell morphology by remodeling the actin cytoskeleton [ 71 ]. The in silico data concerning the possible relationships between lncRNAs and protein-coding genes in porcine granulosa cells treated with TCDD are supported by the results of our recent study [ 72 ]. In this study, the abundance of heat shock proteins as well as cytoskeleton and ECM proteins were significantly affected by TCDD in porcine granulosa cells.…”

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the transcriptome of aryl hydrocarbon receptor (AhR) knock-down porcine granulosa cells

“…The AVG-16 cell line was derived from granulosa cells of medium porcine follicles (Horisberger, 2006), and the cells were reported to exhibit AhR expression, both at mRNA and protein level (Sadowska et al, 2015). Previously, we employed the intact (untransfected) AVG-16 cells to study the impact of TCDD on the gene (Sadowska et al, 2017;Nynca et al, 2019) and lncRNA expression profiles (Ruszkowska et al, 2018) as well as on the cell proteome (Orlowska et al, 2018). The AhR gene and protein abundance in the cultured AhR knock-down AVG-16 cells was significantly reduced in comparison to control untransfected cells.…”

“…In the current study, the expression of all these genes was up-regulated by TCDD. Recently, it was demonstrated in intact porcine granulosa cells that TCDD also affected the abundance of genes (Sadowska et al, 2017) and proteins (Orlowska et al, 2018) involved in cell proliferation and differentiation. These findings suggest that TCDD may disturb the cell cycle progression regardless of the AhR status of the granulosa cells.…”

“…In the present study, we were focused on identifying all possible pathways (molecules) involved in the mechanism of TCDD action in porcine granulosa cells and, therefore, the selected dose of TCDD (100 nM) moderately exceeded its environmentally relevant concentrations. The concentration of 100 nM of TCDD was previously reported to affect porcine granulosa cells steroidogenesis (Gregoraszczuk et al, 2000;Gregoraszczuk, 2002;Jablonska et al, 2014), gene expression (Piasecka-Srader et al, 2016;Sadowska et al, 2017;Nynca et al, 2019), lncRNA expression (Ruszkowska et al, 2018) and proteome (Orlowska et al, 2018) without affecting cell viability and morphology. Moreover, 100 nM was one of the two most frequently used TCDD concentrations in in vitro experiments performed on porcine ovarian cells.…”

“…The AhR abundance was determined in C UT , C NEG and C S cells after 27 (24 h of transfection + 3 h of cell culture), 36 (24 h of transfection + 12 h of cell culture) and 48 (24 h of transfection + 24 h of cell culture) hours of culture. Protein isolation and western blot analysis were performed as we previously described (Orlowska et al, 2018). Protein samples (15 µg) were separated by SDS-PAGE (9% polyacrylamide gel) and transferred to nitrocellulose membranes (GE Healthcare, Little Chalfont, UK) using the Owl TM VEP-2 Mini Tank Electroblotting System (Thermofisher Scientific) for wet electroblotting (90 mA, 70 min).…”