Is cytochrome P450 3A4 regulated by menstrual cycle hormones in control endometrium and endometriosis?

The estrogen-metabolizing activities of cytochrome P450 (CYP) enzymes have been implicated in endometriosis. However, their regulation in various sources of endometrial tissue under different hormonal conditions has not been clarified. Our objective was to study the hormone regulation of a specific CYP enzyme, namely CYP3A4, in control (n = 15) and endometriosis patients (n = 42). To this end, we evaluated mRNA expression (using real-time PCR) of CYP3A4 in tissue samples classified according to the phase of menstrual cycle at which they were obtained as confirmed by the related circulating hormone levels. Protein expression was also evaluated by Western Blot. In order to further investigate the hormonal regulation of CYP3A4, stromal cells from ovarian endometriotic lesions were cultured with the prevailing hormones of the distinct phases of the menstrual cycle. We observed that all control and endometriosis tissues express CYP3A4. Nevertheless, changes in CYP3A4 gene expression related to cycle phase were only seen in the control eutopic endometrium and not in samples from endometriosis patients, with an increase in the luteal phase. Stromal cells isolated from ovarian endometriotic lesions expressed CYP3A4 and their exposure to luteal phase-mimicking hormones (estradiol + progesterone) reduced CYP3A4 mRNA in parallel with a diminished expression of the corresponding receptors, estrogen receptor alpha and progesterone receptor. Our findings suggest that steroid hormones are able to regulate CYP3A4 mRNA expression, although the circulating levels of these hormones can only regulate control endometrium and not endometriosis tissues, probably because of dysregulated local steroid concentration in these latter samples. Similar content being viewed by others

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The authors would like to acknowledge Albina Verceze for hormonal measurements and Gabriela dos S. Hidalgo and Marta Diniz, for technical assistance during sample collection and storage. Thanks to Isis Mozetic for artwork assistance, Prof. Ises de Almeida Abrahamsohn and Prof. Sergio Podgaec for critically reading the manuscript. Special thanks also to Elivane da Silva Victor and Deisy Morselli Victor for statistical consulting. This work was supported by the São Paulo Research Foundation (FAPESP: 2010/02412-6) and CNPq/INCT-Hormones. Author information Authors and Affiliations Corresponding author Electronic supplementary material Below is the link to the electronic supplementary material. 11010_2016_2899_MOESM1_ESM.tif (download TIF ) Supplementary Figure 1. Representative blots of CYP3A4 protein levels in different sites. Protein extract of the same samples were analyzed by Western Blot. Blots of protein expression of CYP3A4 in endometria and lesions are represented. Samples: 1 and 2: control endometrium, 3 and 4: eutopic endometrium, 5: non deep-infiltrating lesion, 7 and 8: deep-infiltrating lesions. (TIFF 1289 kb) 11010_2016_2899_MOESM2_ESM.tif (download TIF ) Supplementary Figure 2. Dose–response of estradiol in the CYP3A4 mRNA expression in stromal cells. The gene expression levels of ESR1, PR and CYP3A4 were analyzed by real time PCR in ovarian endometriotic stromal cells treated with estradiol and vehicle. Relative gene expression levels were normalized by Genorm algorithm and to the vehicle-treated ovarian stromal cells (control). No statistical differences between groups are observed (P > 0.05). (TIFF 4605 kb) Rights and permissions About this article Cite this article Piccinato, C.A., Neme, R.M., Torres, N. et al. Is cytochrome P450 3A4 regulated by menstrual cycle hormones in control endometrium and endometriosis?. Mol Cell Biochem 427, 81–89 (2017). https://doi.org/10.1007/s11010-016-2899-3 Received: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s11010-016-2899-3