phytoestrogens and phorbolol-myristate-acetate

Corpus luteum (CL) is a reproductive gland that plays a crucial endocrine role in the regulation of the estrous cycle, fertility, and pregnancy in cattle. The main function of CL is secretion of progesterone (P4), an important hormone for establishment a successful pregnancy, whereas prostaglandin F(2alpha) (PGF(2alpha)), 17beta-estradiol (E(2)) and testosterone (T) are implicated in the regulation of luteolysis. It has been shown that phytoestrogens may disrupt numerous reproductive functions on several levels of regulation and via different intracellular mechanisms. Using a cell-culture system of steroidogenic cells of the bovine CL, we determined effects of active phytoestrogen metabolites (equol and para-ethyl-phenol) on PGF(2alpha), P4, and T synthesis in steroidogenic CL cells. Moreover, we examined the intracellular mechanisms of phytoestrogen metabolite actions. Phytoestrogen metabolites did not affect P4 production in steroidogenic CL cells. However, PGF(2alpha) and T were significantly stimulated by metabolites of phytoestrogens in the bovine steroidogenic CL cells. To study the intracellular mechanism of endogenous E(2) and phytoestrogen metabolites action, steroidogenic cells were preincubated with a phospholipase C inhibitor (U73122), a protein kinase C inhibitor (staurosporine), an estrogen receptor antagonist (ICI) and a transcription inhibitor (actinomycin D) for 0.5h, and then stimulated with para-ethyl-phenol, equol or E(2). Only U73122 and staurosporine totally reduced the stimulatory effect of E(2) on PGF(2alpha) production by the cells. ICI and actinomycin D only partially reduced E(2) action on CL cells. In contrast, the stimulatory effect of phytoestrogen metabolites was totally inhibited by ICI and actinomycin D. Moreover, in contrast to E(2) action, phytoestrogen metabolites did not cause intracellular calcium mobilization in the cells. The present study demonstrated that phytoestrogen metabolites stimulate PGF(2alpha) secretion in steroidogenic cells of the bovine CL via the estrogen receptor-dependent, genomic pathway.

Topics: Animals; Calcium; Cattle; Cells, Cultured; Corpus Luteum; Dactinomycin; Diet; Dinoprost; Equol; Estrenes; Female; Glycine max; Isoflavones; Luteinizing Hormone; Phenols; Phytoestrogens; Pyrrolidinones; Staurosporine; T-Lymphocytes; Tetradecanoylphorbol Acetate

Epidemiological studies indicate that Asian women have a lower incidence of breast cancer compared with their counterparts in the West, and soya consumption has been suggested as a contributory factor. Clinical and animal studies have revealed that cyclooxygenase-2 (COX-2) expression is associated with a risk of breast cancer. In the present study, we investigated the effect of soya isoflavones on the expression of COX-2 in the breast cell line MCF-7. Genistein, daidzein and equol were found to inhibit COX-2 expression induced by phorbol 12-myristate 13-acetate (PMA). Similar findings were observed in the COX-2 protein analysis. In order to study transcriptional control, a fragment of the 5'-flanking region of the hCOX-2 gene was amplified and inserted into a firefly luciferase reporter plasmid. The reporter assay indicated that the transactivation of the hCOX-2 promoter was induced by PMA, and activity was inhibited with the co-administration of genistein, daidzein or equol. An activator protein-1 (AP-1)/cyclic AMP response element binding protein (CREB) binding site (-59/-53) was identified in hCOX-2 promoter, and this could be critical in PMA-induced COX-2 expression. Truncation reporter plasmids with (-70/-36) and without (-51/-36) AP-1/CREB were constructed for subsequent analysis. The results revealed that the hCOX-2 promoter transactivation suppressed by isoflavone could be dependent on AP-1/CREB binding. Nonetheless, this study illustrated that the soya isoflavones reduced COX-2 expression, which could be important in the post-initiation events of breast carcinogenesis.

Topics: Anticarcinogenic Agents; Breast Neoplasms; Carcinogens; Cell Line, Tumor; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Equol; Female; Genistein; Glycine max; Humans; Isoflavones; Membrane Proteins; Phytoestrogens; Plant Extracts; Promoter Regions, Genetic; Tetradecanoylphorbol Acetate