Effect of TCDD Exposure on CYP1A1 and CYP1B1 Expression in Explant Cultures of Human Endometrium
| Autor: | Thomas R. Sutter, Li Feng, Frank D. Stephen, Kevin G. Osteen, James R. Olson, John Love, Theodore G. Costich, Diane P. Bofinger, Ronald E. Batt, Lai-Har Chi, Stephen T. Koury |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
medicine.medical_specialty
Polychlorinated Dibenzodioxins Uterus Endometriosis Biology Toxicology Endometrium Gene Expression Regulation Enzymologic Andrology Cytochrome P-450 Enzyme System Culture Techniques Internal medicine Gene expression Cytochrome P-450 CYP1A1 medicine Humans Northern blot DNA Primers Base Sequence Reverse Transcriptase Polymerase Chain Reaction Aryl Hydrocarbon Receptor Nuclear Translocator medicine.disease Immunohistochemistry DNA-Binding Proteins Blot medicine.anatomical_structure Endocrinology Receptors Aryl Hydrocarbon Cytochrome P-450 CYP1B1 Female Aryl Hydrocarbon Hydroxylases Transcription Factors Explant culture Hormone |
| Zdroj: | Toxicological Sciences. 62:299-314 |
| ISSN: | 1096-0929 |
| DOI: | 10.1093/toxsci/62.2.299 |
| Popis: | Endometriosis is a debilitating disease estimated to affect 10% of reproductive-age women and characterized by the growth of endometrial tissue outside of the uterus. The present study char- acterizes a human endometrial explant culture model for studying the direct effects of TCDD exposure by assessing the expression of CYP1A1 and CYP1B1 mRNA (Northern blotting), protein (West- ern blotting), and activity (7-ethoxyresorufin-O-deethylase; EROD) in explants cultured with and without TCDD. Explants were obtained at laparoscopy or laparotomy from women under- going surgery for tubal ligation, endometriosis, or pelvic pain unrelated to endometriosis. The explants were cultured with 10 nM estradiol (E2 )o r 1n M E 2 plus 500 nM progesterone (P4) with or without TCDD (first 24 h). The expression of CYP1A1 and CYP1B1 mRNA was greatest with 10 nM TCDD and increased up to 72 h after initial exposure. EROD activity increased up to 120 h. Explants from a secretory phase biopsy became reorganized in culture and formed a new epithelial membrane, while maintaining basic endometrial morphology and viability for up to 120 h. At 24 h, TCDD significantly increased CYP1A1 and CYP1B1 mRNA, and at 72 h, TCDD significantly increased EROD activity and CYP1B1 protein compared to explants cultured without TCDD for similar times. CYP1B1 protein also exhibited substan- tial constitutive expression that was similar in uncultured biopsies, where CYP1B1 protein was immunolocalized in the cytoplasm of epithelial glands, with only occasional patches of protein in the surface epithelial membrane. In explants cultured with and with- out TCDD exposure, CYP1B1 protein was localized in the cyto- plasm of the new surface epithelial membrane and glands closest to the surface. CYP1A1 protein was not detected in uncultured biopsies or explants. Both younger age (age 30 and under) and proliferative phase were associated with higher TCDD-induced EROD activity in specimens treated with E2:P4. No significant endometriosis-related differences were observed for any of the biomarkers, but the detection of disease-specific change was lim- ited by small sample size and variability in tissue-cycle phase. The human endometrial explant culture model will be useful for future studies of the effects of dioxin-like compounds on human endo- metrium in relationship to cycle phase and hormonal exposure. |
| Databáze: | OpenAIRE |
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