Project description:Here we report the gene expression profile of in vitro cultured human endometrial stromal cells treated with siRNA targeting FOXO1 piror to eutherian differentiation media exposure. The eutherian differentiation media contains cyclic AMP (cAMP) analogue 8-Br-cAMP and the progesterone (P4) analogue medroxyprogesterone acetate (MPA).
Project description:Serum starved MDAMB453 cells were serum starved and treated for 4 hours with vehicle control (ethanol), 5α-dihydrotestosterone (DHT; 10nM) or medroxyprogesterone acetate (MPA; 10nM). AR binidng peaks were determined. Subsequent analysis showed that binding sites for androgen receptor are enriched at breast cancer risk loci.
Project description:Comparison between the effects of progesterone (P4) and medroxyprogesterone acetate (MPA) combined to estradiol (E2) on gene expression in normal breast cells to provide insight on their possible different impact on breast cancer risk in vivo.
Project description:Medroxyprogesterone acetate (MPA) is a progestin that can bind to and activate progesterone, androgen and glucocorticoid receptors. However, it is not known which receptor mediates MPA action in a cellular context where all three of these receptors are co-expressed and functional. This microarray experiment was performed to compare the transcriptomes induced by MPA and the cognate ligands for these receptors ie progesterone (PROG), 5a-dihydrotestosterone (DHT) and dexamethasone (DEX) in breast cancer cells to determine which was most similar to MPA.
Project description:Molecular mechanisms responsible for abnormal endometrial vasculature in women receiving long-acting progestin-only contraceptives (LAPCs) are unknown. We hypothesize that LAPCs impairs vascular smooth muscle cell (VSMC) and pericyte proliferation and/or migration producing thin-walled hyperdilated fragile microvessels prone to bleeding. Proliferating cell nuclear antigen (PCNA) and alpha smooth muscle actin (aSMA) double-immunohistochemistry assessed VSMC differentiation and proliferation in endometria from women pre- and postDepoProvera (Depo) treatment and from oophorectomized guinea pigs (OVX-GPs) treated with vehicle, estradiol (E2), medroxyprogesterone acetate (MPA) or E2+MPA pellets. After treating cultured VSMCs with MPA or etonogestrel (ETO), whole genome profiling, proliferation and migration assays were performed. Endometrial vasculature of Depo-administered women displayed reduced M-DM-.SMA immunoreactivity and fewer PCNA (+) nuclei among aSMA (+) cells (P<0.008). Microarray analysis of VSMCs identified several MPA and ETO-altered transcripts regulated by STAT1 signaling (p<2.22x10-6), including chemokine (C-C motif) ligand 2 (CCL2). Both MPA and ETO reduced VSMC proliferation and migration (p<0.001), recombinant CCL2 reversed this progestin inhibition, and, in turn, a STAT1 inhibitor abolished these CCL2 effects. Similarly, the endometria of MPA treated OVX-GPs displayed decreased aSMA staining and fewer PCNA (+) nuclei in VSMC (p<0.005). In conclusion, LAPCs promote abnormal endometrial vessel formation by inhibiting VSMC proliferation and migration. Total RNA (n=3) obtained from cultured TDCs incubated 6h with estradiol or estradio + medroxyprogesterone acetate with or without 1 ng/ml IL-1M-NM-2 for 6h.
Project description:Noval and traditional signaling pathways involved in cervical ripening that were regulated by MPA were identified. Experiment Overall Design: MPA treatment on E15 pregnant cervix (n=6) vs Vehicle group (n=6)
Project description:DNA methyaltion analysis using Illumina Infinium Mouse Methylation Beadchips in different groups of mice with and without exposure to medroxyprogesterone acetate and 7,12-dimethylbenzanthracene MPA/DMBA (which induces mammary gland tumours), with and without mifepristone (an antiprogestin) treatment, and with and without a ketogenic diet.
Project description:Comparison between the effects of progesterone (P4) and medroxyprogesterone acetate (MPA) combined to estradiol (E2) on gene expression in normal breast cells to provide insight on their possible different impact on breast cancer risk in vivo. Total RNA obtained from cultures of normal human breast epithelial cells (HBE) under E2, E2+MPA and E2+P4 six hour treatments, compared to untreated HBE cells
Project description:This experiment continues our investigation of glycosylation in endometrial epithelium. During the menstrual cycle, estrogen and progesterone sensitise the uterus to the implanting embryo. There is a major expansion of Golgi and secretory function with increases in mucin (MUC1) and the appearance of specific glycans. We are using Ishikawa cells as a model for in vitro implantation, and have shown the presence of steroid receptors and alterations in glycosylation after hormone exposure. We are using Ishikawa cells as a model for in vitro implantation, and have shown the presence of steroid receptors and alterations in glycosylation after hormone exposure. We now wish to profile glycogenes in steroid (E+P) exposed and control cells. We now wish to profile glycogenes in steroid (E+P) exposed and control cells. Steroid conditions are: estradiol (E2 at 10exp -8M) and medroxyprogesterone acetate (MPA at 10 exp -7M)): 24h with E2, followed by E2+MPA for 72h. Cells were grown on plastic or glass.
