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The role of PPARγ in TBBPA-mediated endocrine disrupting effects in human choriocarcinoma JEG-3 cells.

Honkisz E, Wójtowicz AK - Mol. Cell. Biochem. (2015)

Bottom Line: Our results showed that after TBBPA treatment at 10 nM and 10 µM, PPARγ protein expression increased in a time-dependent manner until 48 h and then slightly decreased at 72 h but was still above the control level.Finally, in the present study, we demonstrated that TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control.These results showed the up-regulation of PPARγ protein expression after TBBPA exposure in human placental cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biotechnology, University of Agriculture, Rędzina 1B, 30-248, Kraków, Poland.

ABSTRACT
The goal of the present study was to investigate the action of TBBPA on PPARγ protein expression in vitro in human choriocarcinoma-derived placental JEG-3 cells. We also analyzed TBBPA for its action on placental secretion of progesterone and β-hCG, cell viability, and apoptosis. Our results showed that after TBBPA treatment at 10 nM and 10 µM, PPARγ protein expression increased in a time-dependent manner until 48 h and then slightly decreased at 72 h but was still above the control level. This alteration in PPARγ protein expression was accompanied by a decreased β-hCG level. Interestingly, co-treatment with the PPARγ antagonist GW9662 reversed the TBBPA-mediated changes in PPARγ protein expression but, according to β-hCG secretion, potentiated an inhibitory effect of TBBPA. Additionally, in our study, we assessed the ability of TBBPA to increase progesterone levels in JEG-3 cells compared with those of controls. Finally, in the present study, we demonstrated that TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control. The apoptotic action of TBBPA was also confirmed by Hoechst 33342 staining. These results showed the up-regulation of PPARγ protein expression after TBBPA exposure in human placental cells. Although co-treatment with antagonist of PPARγ reversed the TBBPA-mediated increase in this protein expression and restored it to the control level, it did not reverse the effect on β-hCG secretion. This indicated that the mechanism of TBBPA-induced changes in β-hCG secretion is PPARγ-independent.

No MeSH data available.


Related in: MedlinePlus

Time- and dose-dependent effect of increasing concentrations of TBBPA on apoptosis in JEG-3 cells as determined by caspase-3 activity. Cells treated with 1 nM of staurosporine (STA) were used as a positive control. Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with ***p < 0.001 reflects statistically significant differences between control and experimental groups
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Fig2: Time- and dose-dependent effect of increasing concentrations of TBBPA on apoptosis in JEG-3 cells as determined by caspase-3 activity. Cells treated with 1 nM of staurosporine (STA) were used as a positive control. Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with ***p < 0.001 reflects statistically significant differences between control and experimental groups

Mentions: TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control (Fig. 2). The caspase-3 activity was dose-dependent up to 10 and 1 µM of TBBPA for 24 and 48 h, respectively. At these concentrations, the average increase in caspase-3 activity was 1.8- to 2.5-fold after 48 h compared with that observed after 24 h. After 72 h exposure to TBBPA, caspase-3 activity reached the level compared to that of the control cells, suggesting that the final phase of apoptosis occurs upon damage to the cell membrane. The apoptotic action of TBBPA was also confirmed by Hoechst 33342 staining (Fig. 3a–d). Morphological changes, as indicated by bright blue fragmented nuclei, nuclear shrinkage, and subsequent chromatin condensation into the periphery of the nuclei, were identified as apoptotic bodies. The number of apoptotic body increased in a dose-dependent manner.Fig. 2


The role of PPARγ in TBBPA-mediated endocrine disrupting effects in human choriocarcinoma JEG-3 cells.

Honkisz E, Wójtowicz AK - Mol. Cell. Biochem. (2015)

Time- and dose-dependent effect of increasing concentrations of TBBPA on apoptosis in JEG-3 cells as determined by caspase-3 activity. Cells treated with 1 nM of staurosporine (STA) were used as a positive control. Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with ***p < 0.001 reflects statistically significant differences between control and experimental groups
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4589557&req=5

Fig2: Time- and dose-dependent effect of increasing concentrations of TBBPA on apoptosis in JEG-3 cells as determined by caspase-3 activity. Cells treated with 1 nM of staurosporine (STA) were used as a positive control. Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with ***p < 0.001 reflects statistically significant differences between control and experimental groups
Mentions: TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control (Fig. 2). The caspase-3 activity was dose-dependent up to 10 and 1 µM of TBBPA for 24 and 48 h, respectively. At these concentrations, the average increase in caspase-3 activity was 1.8- to 2.5-fold after 48 h compared with that observed after 24 h. After 72 h exposure to TBBPA, caspase-3 activity reached the level compared to that of the control cells, suggesting that the final phase of apoptosis occurs upon damage to the cell membrane. The apoptotic action of TBBPA was also confirmed by Hoechst 33342 staining (Fig. 3a–d). Morphological changes, as indicated by bright blue fragmented nuclei, nuclear shrinkage, and subsequent chromatin condensation into the periphery of the nuclei, were identified as apoptotic bodies. The number of apoptotic body increased in a dose-dependent manner.Fig. 2

Bottom Line: Our results showed that after TBBPA treatment at 10 nM and 10 µM, PPARγ protein expression increased in a time-dependent manner until 48 h and then slightly decreased at 72 h but was still above the control level.Finally, in the present study, we demonstrated that TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control.These results showed the up-regulation of PPARγ protein expression after TBBPA exposure in human placental cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biotechnology, University of Agriculture, Rędzina 1B, 30-248, Kraków, Poland.

ABSTRACT
The goal of the present study was to investigate the action of TBBPA on PPARγ protein expression in vitro in human choriocarcinoma-derived placental JEG-3 cells. We also analyzed TBBPA for its action on placental secretion of progesterone and β-hCG, cell viability, and apoptosis. Our results showed that after TBBPA treatment at 10 nM and 10 µM, PPARγ protein expression increased in a time-dependent manner until 48 h and then slightly decreased at 72 h but was still above the control level. This alteration in PPARγ protein expression was accompanied by a decreased β-hCG level. Interestingly, co-treatment with the PPARγ antagonist GW9662 reversed the TBBPA-mediated changes in PPARγ protein expression but, according to β-hCG secretion, potentiated an inhibitory effect of TBBPA. Additionally, in our study, we assessed the ability of TBBPA to increase progesterone levels in JEG-3 cells compared with those of controls. Finally, in the present study, we demonstrated that TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control. The apoptotic action of TBBPA was also confirmed by Hoechst 33342 staining. These results showed the up-regulation of PPARγ protein expression after TBBPA exposure in human placental cells. Although co-treatment with antagonist of PPARγ reversed the TBBPA-mediated increase in this protein expression and restored it to the control level, it did not reverse the effect on β-hCG secretion. This indicated that the mechanism of TBBPA-induced changes in β-hCG secretion is PPARγ-independent.

No MeSH data available.


Related in: MedlinePlus