{"paper_id":"bbe0b4aa-ec35-4c7a-a974-b0efca3737cd","body_text":"Abstract\nThe 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.\nSimilar content being viewed by others\nReferences\nGuengerich FP (1999) Cytochrome P-450 3A4: regulation and role in drug metabolism. Annu Rev Pharmacol Toxicol 39:1–17. doi:10.1146/annurev.pharmtox.39.1.1\nStresser DM, Kupfer D (1998) Prosubstrates of CYP3A4, the major human hepatic cytochrome P450: transformation into substrates by other P450 isoforms. Biochem Pharmacol 55:1861–1871\nYamazaki H, Shimada T (1997) Progesterone and testosterone hydroxylation by cytochromes P450 2C19, 2C9, and 3A4 in human liver microsomes. 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This work was supported by the São Paulo Research Foundation (FAPESP: 2010/02412-6) and CNPq/INCT-Hormones.\nAuthor information\nAuthors and Affiliations\nCorresponding author\nElectronic supplementary material\nBelow is the link to the electronic supplementary material.\n11010_2016_2899_MOESM1_ESM.tif (download TIF )\nSupplementary 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)\n11010_2016_2899_MOESM2_ESM.tif (download TIF )\nSupplementary 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)\nRights and permissions\nAbout this article\nCite this article\nPiccinato, 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\nReceived:\nAccepted:\nPublished:\nIssue date:\nDOI: https://doi.org/10.1007/s11010-016-2899-3","source_license":"CC0","license_restricted":false}