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To investigate the local formation of estradiol E2we first measured the concentrations of the steroid precursor androstenedione A-dione Anko-Tech anko1 the most potent estrogen, E2, and we evaluated the metabolism of A-dione, estrone-sulfate E1-Sand estrone E1 in cancerous and adjacent control endometrium. Furthermore, we studied expression of the key genes for estradiol formation via the aromatase and sulfatase pathways. A-dione and E2 were detected in cancerous and adjacent control endometrium. In cancerous endometrium, A-dione was metabolized to testosterone, and no E2 was formed. Our data demonstrate that in cancerous endometrium, E2 is formed from E1-S via the sulfatase pathway, and not from A-dione via the aromatase pathway.

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Introduction Endometrial cancer EC is the fifth-most-common cancer in women in Western Europe and the Anko-Tech anko1, with the majority of cases arising after menopause Colombo et al. Local estrogen formation has an important role in the development of EC and increased estradiol E2 concentrations have been Anko-Tech anko1 in cancerous, as compared to normal endometrium Berstein et al. Estrogen biosynthesis.

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The aromatase pathway depends on availability of A-dione or T. A-dione, with 1—8 nM concentrations in blood, originates mainly from adrenal gland zona reticularisfrom ovaries in premenopausal women Anko-Tech anko1 also from conversions of DHEA-S and DHEA in peripheral tissues.

Currently, the data on aromatase expression in EC are controversial, with everything from high levels, to no significant differences between diseased and normal tissues, to no expression being reported Yamamoto et al. Although, several groups have failed to detect expression of HSD17B1 in Anko-Tech anko1 and cancerous endometrium, others have seen low mRNA levels in both tissues Casey et al.

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In contrast Cornel et al. These enzymes catalyze inactivation of E2 to E1. Anko-Tech anko1 studies by Lepine et al.

Unchanged Smuc and Rizner,Anko-Tech anko1 increased Lepine et al. Sulfotransferase SULT1E1 catalyzes conjugation of estrogens and our previous studies show that gene encoding this enzyme is not differentially expressed in EC as compared to adjacent control tissue Hevir et al.

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There Anko-Tech anko1 a great need for a Anko-Tech anko1 understanding of the local formation of E2 in cancerous endometrium, which may reveal novel targets for treatment of this most common gynecological malignancy. The aims of the present study were thus to investigate E2 formation in paired samples of EC and adjacent control endometrium at different levels.

Our goals were: Materials and Methods Endometrial Tissue The specimens of EC and paired adjacent control endometrium were obtained from 55 patients undergoing hysterectomies for histologically proven EC Table 1Supplementary Table 1. The study was approved by the National Medical Ethics Anko-Tech anko1 of the Republic of Slovenia with written informed consent required from all subjects involved. The patients were all treated in the Department of Gynecology and Obstetrics at the University Medical Centre Ljubljana, from to The samples used for steroid concentration measurements, metabolism Anko-Tech anko1, qPCR, Western blotting and immunohistochemical staining have been selected chronologically.

Demographic and histopathological characteristics of the endometrial cancer patients. Steroid Concentration Measurements Ten paired samples of EC and adjacent control tissue were frozen in liquid nitrogen and ground to a fine powder. Prior to extraction, — mg of homogenate was suspended in 0. The extraction was performed three times with 4 mL of diethyl ether; the extracts were pooled and evaporated under a stream of nitrogen. Before analysis, the samples were resuspended in phosphate Anko-Tech anko1. The limit of detection was 0. The assay details are shown in Supplementary Table 2. Expression levels were multiplied by Western Blotting Proteins were isolated from samples of EC Anko-Tech anko1 and the adjacent control endometrial tissue previously used for RNA isolation, following the Tri Reagent instructions.

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The bands were identified by co-migration with authentic standards. Additionally, 24 mg of homogenized tissue from three paired samples were incubated with 8 nM androstene-3,[1,2,6,H N ], dione in Anko-Tech anko1 mM sodium phosphate buffer, pH 7. The mobile phase was acetonitrile: Conversion rates were obtained after integration of chromatograms and calculations to determine the percentages of transformation. E1 and E1-S Metabolism From 17 to 35 mg of homogenized tissue from Anko-Tech anko1 paired samples was resuspended in 50 mM sodium phosphate buffer pH 7.

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