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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 JEG-3 cells viability (line graph) and progesterone secretion (bar graph). Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with *P < 0.05;***p < 0.001 (progesterone secretion) and ###p < 0.001 (LDH release) reflects statistically significant differences between control and experimental groups
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Fig1: Time- and dose-dependent effect of increasing concentrations of TBBPA on JEG-3 cells viability (line graph) and progesterone secretion (bar graph). Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with *P < 0.05;***p < 0.001 (progesterone secretion) and ###p < 0.001 (LDH release) reflects statistically significant differences between control and experimental groups

Mentions: Increasing concentrations of TBBPA in the cultures of JEG-3 cells affected progesterone secretion (Fig. 1). TBBPA treatment for 24 h led to an increase in progesterone levels but was only significant in the micromolar range (10 and 50 µM). However, 48 h of treatment with both nanomolar and micromolar doses resulted in significant increases, with progesterone secretion ranging from approximately 12–30 %. TBBPA treatment with 50 µM seemed to be the most effective at increasing progesterone levels by 2- to 3-fold compared with those of the vehicle control (DMSO) at all of the time points. TBBPA at a concentration of 100 µM decreased progesterone secretion due to the dominant cytotoxic effect (up to 53 % LDH leakage).Fig. 1


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 JEG-3 cells viability (line graph) and progesterone secretion (bar graph). Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with *P < 0.05;***p < 0.001 (progesterone secretion) and ###p < 0.001 (LDH release) 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

Fig1: Time- and dose-dependent effect of increasing concentrations of TBBPA on JEG-3 cells viability (line graph) and progesterone secretion (bar graph). Each point represents the mean ± SEM of three independent experiments, each of which consisted of ten replicates per treatment group. Data indicated with *P < 0.05;***p < 0.001 (progesterone secretion) and ###p < 0.001 (LDH release) reflects statistically significant differences between control and experimental groups
Mentions: Increasing concentrations of TBBPA in the cultures of JEG-3 cells affected progesterone secretion (Fig. 1). TBBPA treatment for 24 h led to an increase in progesterone levels but was only significant in the micromolar range (10 and 50 µM). However, 48 h of treatment with both nanomolar and micromolar doses resulted in significant increases, with progesterone secretion ranging from approximately 12–30 %. TBBPA treatment with 50 µM seemed to be the most effective at increasing progesterone levels by 2- to 3-fold compared with those of the vehicle control (DMSO) at all of the time points. TBBPA at a concentration of 100 µM decreased progesterone secretion due to the dominant cytotoxic effect (up to 53 % LDH leakage).Fig. 1

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