The regulation of steroidogenic acute regulatory protein (StAR) gene expression and the synthesis of steroids from cholesterol in ectopic endometriosis tissues were investigated. Peritoneal fluid and endometrial tissues were collected from patients with endometriosis and otherwise healthy women. Peritoneal progesterone and 17 beta-E2 concentrations were highest in early stage endometriosis compared with those in advanced stage endometriosis and in normal women. In concordance with the profile of peritoneal steroids, StAR mRNA and protein were greatest in ectopic implants of early endometriosis. In the advanced stage, concentrations of StAR mRNA and protein were also greater compared with those in normal endometrium. In contrast, P450 side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase transcripts were not different between normal endometrium and ectopic endometriotic implants. Expression of StAR mRNA was detected in purified stromal, but not epithelial, cells. Treatment with PGE(2), but not TNF alpha, or IL-1 beta significantly increased StAR expression and thus induced progesterone production in cultured endometriotic stromal cells. These results demonstrated that aberrant expression of StAR in ectopic endometriotic tissues leading to increased peritoneal progesterone is associated with the formation of endometriosis. Induction of StAR gene expression by peritoneal PGE(2) in endometriotic stromal cells may further contribute to the development of endometriosis.

Fibroblast growth factor-9 (FGF-9) is an autocrine/paracrine growth factor considered to be important for the growth and survival of motorneurons and prostate. In this study, we found that FGF-9 was expressed at high levels in normal uterine endometrium, especially during the late proliferative phase, which is coincident with the rise of estradiol and the time of uterine endometrial proliferation. Using quantitative RT-PCR analysis, we found that FGF-9 mRNA was expressed primarily by endometrial stromal cells. High affinity receptors of FGF-9 were detected in both epithelial and stromal cells but with distinct patterns. FGFR2IIIc and FGFR3IIIc are abundant in endometrial stromal cell. FGFR2IIIb is mostly expressed in endometrial epithelial cells, whereas FGFR3IIIb is found in both epithelial and stromal cells. Treatment with FGF-9 induces endometrial stromal proliferation in a dose-dependent manner. Expression of FGF-9 in stromal cells was induced by 17beta-estradiol but not by progesterone. Furthermore, the administration of 17beta-estradiol stimulates endometrial stromal cell proliferation and that can be inhibited by cotreatment with anti-FGF-9 antibody. Herein we demonstrate, for the first time, that FGF-9 is an autocrine estromedin endometrial stromal growth factor that plays roles in cyclic proliferation of uterine endometrial stroma.

Fibroblast growth factor-9 (FGF-9) is a steroid-regulated mitogen and survival factor for nerve and mesenchymal cells. In the current study, we determined the expression pattern and functional roles of FGF-9 in the ectopic endometriotic lesions. We found that FGF-9 and its receptors were effectively expressed by ectopic endometriotic tissues. The expression of FGF-9 was greater in the early stage of endometriosis, compared with the severe stage, which is consistent with concentration of 17 beta-estradiol in the peritoneal fluid of women with endometriosis. In addition, expression of FGF-9 in ectopic endometriotic stromal cell was inhibited by treatment with ICI 182,870 indicating it is likely regulated by estrogen in an autocrine manner. Administration of 17 beta-estradiol induced FGF-9, FGF receptor 2IIIc, and FGF receptor 3IIIc expression in endometriotic stromal cells. Concordant with this result, treatment of endometriotic stromal cells with 4-hydroxyandrostenedione (an aromatase inhibitor) or ICI 182,870 inhibited their proliferation, and that was reversed by coadministration with 17 beta-estradiol or FGF-9. In conclusion, expression of FGF-9 in endometriotic stromal cells is associated with aberrant production of estrogen. The capability of proliferation possessed by endometriotic stromal cell during menstruation when ovarian 17 beta-estradiol is in the nadir may be mediated, at least in part, by autocrined estrogen-stimulated expression of FGF-9 and its receptors.

A hydrophilic matrix of periodate-oxidized dextran was used as a double-sided linker to covalently immobilize Staphylococcus aureus protein A (SpA) molecules onto a poly-L-lysine-modified piezoelectric crystal surface to improve their stability, activity, and binding specificity with human immunoglobulin G (IgG) in flow injection assays. The prepared sensing crystals displayed best sensitivity and reusability at a flow rate of 140 microL/min. A human IgG concentration as low as 0.3 nM can be detected by this system. Up to 19 successive assay repetitions were achieved without significant loss of sensitivity using the same crystal. The analysis of adsorption kinetics indicates that such a preparation can greatly increase the amount of available active human IgG binding sites on immobilized SpA. Hardly any response arising from unspecific binding was detected. In addition, the sensing crystal prepared by this method was found to retain activity better than one prepared via direct deposition when stored in either wet or dry states. Finally, the prepared SpA-coated crystals were applied to the affinity immobilization of polyclonal goat anti-Schistosoma japonicum glutathione-S-transferase (GST) and were able to subsequently detect GST and its genetically engineered mutant either in a purified form or in the crude cell lysate.

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