The effects of prochloraz on the levels of nuclear receptor genes expressions and global DNA methylation in human prostate carcinoma cells

Abstract

Background and Aims: Prochloraz (PCZ) is an imidazole fungicide which is used in agriculture and gardening. PCZ, with endocrine disrupting effect, disrupts reproductive and developmental functions. Previously, the effects of PCZ on the estrogen and androgen receptors have been shown in vitro and in vivo. Because of endocrine disrupting effects, PCZ could influence nuclear receptors which acted as ER and AR antagonists. Besides, PCZ has been thought to have no genotoxic effects. Therefore, we aimed to investigate possible effects of PCZ on nuclear receptor genes and epigenetic mechanisms in human prostate carcinoma (PC-3) cells. Methods: In the present study, MTT and LDH tests were applied to evaluate the cell viability. Expression levels of nuclear receptor genes such as AhR, PXR, PPARα, PPARγ were studied on real-time quantitative PCR. For global DNA methylation analysis, the levels of 5-methylcytosine (5-mC%) were measured by elisa kit. Results: According to MTT and LDH test results, IC50 value of PCZ has been determined as 144.19 and 116.65 μM, respectively. There were significant changes for the expression levels of AhR, PPARα and PPARγ genes after 5-50 μM of PCZ treatments. 5 and 50 μM of PCZ decreased the levels of 5-mC% in the rates of 22.6% and 26.9%, respectively. Conclusion: It has been suggested that PCZ may cause alterations on the expressions of nuclear receptor genes which could be related to endocrine disrupting effects and may have implications on global DNA methylation.

Keywords

• Cytotoxicity
• DNA methylation
• nuclear receptor genes
• PC-3 cells
• prochloraz

Supporting Institution

Scientific Research Projects Coordination Unit of Istanbul University

Project Number

TLO-2017-23409; TLO-2017-27475

Thanks

This work was supported in part by the Scientific Research Projects Coordination Unit of Istanbul University (Project numbers: TLO-2017-23409; TLO-2017-27475) and TUBITAK-BIDEB (Project numbers: 2211-C/2015-2 Grant for Ecem Fatma Karaman).

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