Introduction/Background Epidemiological studies have shown that the human follicular fluid (FF) include endocrine-disrupting chemicals (EDCs) such as perfluorooctanoate (PFOA) perfluorooctane sulfonate (PFOS), 2,2-dichlorodiphenyldichloroethylene (p,p’-DDE), polychlorinated biphenyl 153 (PCB153) and hexachlorobenzene (HCB).^{1 2} Chemicals accumulated in FF may exert their potential effects on granulosa cell tumors (GCTs) that arise from the granulosa cells. Two subtypes of GCTs have been described: the juvenile (JGCT) and the adult (AGCT) form.³ GCTs are hormonally active and produce of 17β-estradiol (E2).⁴ Therefore, the aim of this research is to determine whether chemicals with endocrine potential present in FF disrupt E2 secretion by JGCT and AGCT.

Methodology Two human GCT-derived cell lines: COV434 (ECACC) representing JGCT and KGN (Riken Cell Bank) representing AGCT were cultured in three-dimensional culture. To form spheroids, 6000 cells were seeded in 96-well plates. Spheroids were exposed to single chemicals: PFOA (2 ng/ml), PFOS (8 ng/ml), DDE (1 ng/ml), HCB (50 pg/ml) and PCB153 (100 pg/ml) or the mixture of these compounds. The CYP19A1 and CYP1A1 expression were determined by Real-Time PCR analysis. The E2, 2-hydroxyestrogen1 (2-OHE1) and 16-hydroxyestrogen1 (16-OHE1) secretion were measured by ELISA.

Results Firstly, we founded that both basal and T-induced E2 secretion as well as CYP19A1 expression was significantly lower in COV434 than KGN cells. Interestingly, estrogen metabolism potential measured as CYP1A1 expression was contrary correlated. In COV434 cell line, all compounds present in FF and mixtures significantly decreased T-stimulated E2 secretion and simultaneously they increased 2-OHE1/E2 and 16-OHE1/E2 ratio. However, in KGN cells, all compounds and mixtures significantly increased T-stimulated E2 secretion and reduced 2-OHE1/E2 and 16-OHE1/E2 ratio.

Conclusion These results indicated that EDCs action on E2 secretion depends on estrogen metabolism potential of the target tissue. In JGCT which possess higher estrogen metabolism potential, EDCs accumulated in FF stimulated E2 metabolism. However, in AGCT with lower estrogen metabolism potential, EDCs stimulated E2 secretion.

Disclosure This study was funded by the National Science Centre (NCN) Poland (grant number 2016/21/B/NZ7/01080).

References

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2. Petro EM, D’Hollander W, Covaci A, et al. Sci Total Environ. 2014;496:2828.

3. Pectasides D, Papaxoinis G, Fountzilas G, et al. Anticancer Res. 2008;28:1421.

4. Bulun SE, Simpson ER. Adv Exp Med Biol. 2008;630:112–132.