SFRP1 reduction results in an increased sensitivity to TGF-ß signaling.

BACKGROUND: Transforming growth factor (TGF)-ß plays a dual role during mammary gland development and tumorigenesis and has been shown to stimulate epithelial-mesenchymal transition (EMT) as well as cellular migration. The Wnt/ß-catenin pathway is also implicated in EMT and inappropriate activation of the Wnt/ß-catenin signaling pathway leads to the development of several human cancers, including breast cancer. Secreted frizzled-related protein 1 (SFRP1) antagonizes this pathway and loss of SFRP1 expression is frequently observed in breast tumors and breast cancer cell lines. We previously showed that when SFRP1 is knocked down in immortalized non-malignant mammary epithelial cells, the cells (TERT-siSFRP1) acquire characteristics associated with breast tumor initiating cells. The phenotypic and genotypic changes that occur in response to SFRP1 loss are consistent with EMT, including a substantial increase in the expression of ZEB2. Considering that ZEB2 has been shown to interact with mediators of TGF-ß signaling, we sought to determine whether TGF-ß signaling is altered in TERT-siSFRP1 cells. METHODS: Luciferase reporter assays and real-time PCR analysis were employed to measure TGF-ß transcriptional targets. Western blot analysis was used to evaluate TGF-ß-mediated ERK1/2 phosphorylation. Migration chamber assays were utilized to quantify cellular migration. TERT-siSFRP1 cells were transfected with Stealth RNAi™ siRNA in order to knock-down the expression of ZEB2. RESULTS: TERT-siSFRP1 cells exhibit a significant increase in both TGF-ß-mediated luciferase activity as well as TGF-ß transcriptional targets, including Integrin ß3 and PAI-1. Phosphorylation of ERK1/2 is increased in TERT-siSFRP1 cells in response to enhanced TGF-ß signaling. Furthermore, when the TGF-ß pathway is blocked with a TGF-ßR antagonist (LY364947), cellular migration is significantly hindered. Finally, we found that when ZEB2 is knocked-down, there is a significant reduction in the expression of exogeneous and endogenous TGF-ß transcriptional targets and cellular migration is impeded. CONCLUSIONS: We demonstrate that down-regulation of SFRP1 renders mammary epithelial cells more sensitive to TGF-ß signaling which can be partially ameliorated by blocking the expression of ZEB2.

Anxiety disorders during pregnancy: a systematic review.

OBJECTIVE: To systematically evaluate the literature on anxiety disorders during pregnancy. DATA SOURCES: MEDLINE, PsycINFO, and CINAHL were searched through October 2013 for original research studies published in English using combinations of the terms pregnancy, prenatal, or pregnancy outcomes; anxiety disorder; and generalized anxiety. Reference lists of included studies were hand-searched and a PubMed search for in-process reports was conducted. STUDY SELECTION: Relevant studies of anxiety disorders during pregnancy as determined by diagnostic interview were included if they reported on prevalence; course, onset, and/or risk factors; maternal, obstetric, or fetal/child outcomes; and/or treatment trial results. DATA EXTRACTION: Two reviewers independently extracted relevant data and assessed methodological quality of each study. RESULTS: Fifty-seven reports were included. Reports provided information on panic disorder (25 reports), generalized anxiety disorder (17 reports), obsessive-compulsive disorder (OCD) (23 reports), agoraphobia (6 reports), specific phobia (10 reports), social phobia (14 reports), posttraumatic stress disorder (14 reports), and any anxiety disorder (18 reports). Twenty reports provided information on prevalence, 16 on course, 10 on risk factors, and 22 on outcomes. Only 1 treatment study was identified. High anxiety disorder prevalence in pregnancy was found; however, estimates vary considerably, and evidence is inconclusive as to whether prevalence among pregnant women differs from that of nonpregnant populations. Considerable variation in prenatal course of OCD and panic disorder was found. Substantial heterogeneity limits conclusions regarding risk factors or outcomes. CONCLUSIONS: Additional research of higher methodological quality is required to more accurately determine prevalence, understand course, identify risk factors and outcomes, and determine effective treatments for anxiety disorders in pregnancy.