Association of WT1 and induction of apoptosis in endometriotic cell lines treated with MIS

Abstract

OBJECTIVE: Products of steroidogenic factor-1 (SF-1) and Wilms' tumor-1(WT1) genes are essential for mammalian gonadogenesis prior to sexual differentiation. Prior studies demonstrated WT1 inhibits cAMP SF-1 pathway dependent p450arom expression in cultured endometriotic and endometrial stromal cells. Mullerian inhibiting substance is first identified as a factor that causes regression of Mullerian ducts in developing male embryos. Recent research has revealed MIS causes cell-cycle arrest in ovarian, cervical and endometrial cancer cell lines in vitro. In our unpublished data, we showed MIS induces apoptosis and inhibits basal autophagy in ectopic endometrial cells. We hypothesized that WT1 and cAMP SF-1 dependent p450arom plays a role in the effect of MIS on endometriotic cell lines. In this study we aimed to explain the association of WT1 and induction of apoptosis with MIS treatment in endometriosis cell lines.DESIGN: We studied WT1 protein expression in endometriotic cell lines treated with MIS compared to no treatment.MATERIALS AND METHODS: Cultured CRL-7566 endometriosis cell lines from ovarian cyst (ATCC) were harvested at 80% confluency and placed in wells. Each well was treated with 5mcg/ml human recombinant MIS twice 4 days apart or with PBS as control. WT1 protein expression was determined using Western analysis and normalized to β-actin.RESULTS: MIS treatment significantly induced WT1 expression on endometriotic stromal cells(p<0.05) compared to the sample treated with control.CONCLUSION: MIS induces apoptosis, inhibits cellular growth and basal autophagy in endometriosis cell lines. MIS treatment induces WT1 expression which may partially explain the pathway that leads to apoptosis. OBJECTIVE: Products of steroidogenic factor-1 (SF-1) and Wilms' tumor-1(WT1) genes are essential for mammalian gonadogenesis prior to sexual differentiation. Prior studies demonstrated WT1 inhibits cAMP SF-1 pathway dependent p450arom expression in cultured endometriotic and endometrial stromal cells. Mullerian inhibiting substance is first identified as a factor that causes regression of Mullerian ducts in developing male embryos. Recent research has revealed MIS causes cell-cycle arrest in ovarian, cervical and endometrial cancer cell lines in vitro. In our unpublished data, we showed MIS induces apoptosis and inhibits basal autophagy in ectopic endometrial cells. We hypothesized that WT1 and cAMP SF-1 dependent p450arom plays a role in the effect of MIS on endometriotic cell lines. In this study we aimed to explain the association of WT1 and induction of apoptosis with MIS treatment in endometriosis cell lines. DESIGN: We studied WT1 protein expression in endometriotic cell lines treated with MIS compared to no treatment. MATERIALS AND METHODS: Cultured CRL-7566 endometriosis cell lines from ovarian cyst (ATCC) were harvested at 80% confluency and placed in wells. Each well was treated with 5mcg/ml human recombinant MIS twice 4 days apart or with PBS as control. WT1 protein expression was determined using Western analysis and normalized to β-actin. RESULTS: MIS treatment significantly induced WT1 expression on endometriotic stromal cells(p<0.05) compared to the sample treated with control. CONCLUSION: MIS induces apoptosis, inhibits cellular growth and basal autophagy in endometriosis cell lines. MIS treatment induces WT1 expression which may partially explain the pathway that leads to apoptosis.