Jean-Francois Mouillet1, Takuya Mishima1, Andrea Mollica do Amarante Paffaro2, Tony W Parks3, Judy A Ziegler4, Tianjiao Chu1, Yoel Sadovsky5. 1. Magee-Womens Research Institute, University of Pittsburgh, PA, USA; Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, PA, USA. 2. Magee-Womens Research Institute, University of Pittsburgh, PA, USA; Instituto de Ciencias Biologicas, Universidade Federal de Alfenas (UNIFAL-MG), Alfenas, MG, Brazil. 3. Magee-Womens Research Institute, University of Pittsburgh, PA, USA; Department of Pathology, University of Pittsburgh, PA, USA. 4. Magee-Womens Research Institute, University of Pittsburgh, PA, USA. 5. Magee-Womens Research Institute, University of Pittsburgh, PA, USA; Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, PA, USA; Department of Microbiology and Molecular Genetics, University of Pittsburgh, PA, USA. Electronic address: ysadovsky@mwri.magee.edu.
Abstract
INTRODUCTION: Follistatin-like-1 (FSTL1) is a widely expressed secreted protein with diverse but poorly understood functions. Originally described as a pro-inflammatory molecule, it has recently been reported to play a role in signaling pathways that regulate development and homeostasis. Distinctively, FSTL1 harbors within its 3'-UTR the sequence encoding microRNA-198 (miR-198), shown to be inversely regulated relative to FSTL1 expression and to exhibit opposite actions on cellular processes such as cell migration. We sought to investigate the expression of FSTL1 and to assess its interplay with miR-198 in human trophoblasts. METHODS: We used a combination of northern blot analyses, quantitative PCR, small RNA sequencing, western blot and immunohistochemistry to characterize FSTL1 and miR-198 expression in placental trophoblasts. We also used reporter assays to examine the post-transcriptional regulation of FSTL1 and assess its putative regulation by miR-198. RESULTS: We detected the expression of FSTL1 transcript in both the human extravillous trophoblast line HTR-8/SVneo and in primary term human villous trophoblasts. We also found that the expression of FSTL1 was largely restricted to extravillous trophoblasts. Hypoxia enhanced the expression of FSTL1 protein in cultured primary villous trophoblasts. Interestingly, we did not detect any evidence for expression or function of mature miR-198 in human trophoblasts. DISCUSSION: Our data indicate that placental FSTL1 is expressed particularly in extravillous trophoblasts. We also found no evidence for placental expression of miR-198, or for its regulation of FSTL1, implying that the post-transcriptional regulation of FSTL1 by miR-198 is tissue specific.
INTRODUCTION:Follistatin-like-1 (FSTL1) is a widely expressed secreted protein with diverse but poorly understood functions. Originally described as a pro-inflammatory molecule, it has recently been reported to play a role in signaling pathways that regulate development and homeostasis. Distinctively, FSTL1 harbors within its 3'-UTR the sequence encoding microRNA-198 (miR-198), shown to be inversely regulated relative to FSTL1expression and to exhibit opposite actions on cellular processes such as cell migration. We sought to investigate the expression of FSTL1 and to assess its interplay with miR-198 in human trophoblasts. METHODS: We used a combination of northern blot analyses, quantitative PCR, small RNA sequencing, western blot and immunohistochemistry to characterize FSTL1 and miR-198expression in placental trophoblasts. We also used reporter assays to examine the post-transcriptional regulation of FSTL1 and assess its putative regulation by miR-198. RESULTS: We detected the expression of FSTL1 transcript in both the human extravillous trophoblast line HTR-8/SVneo and in primary term human villous trophoblasts. We also found that the expression of FSTL1 was largely restricted to extravillous trophoblasts. Hypoxia enhanced the expression of FSTL1 protein in cultured primary villous trophoblasts. Interestingly, we did not detect any evidence for expression or function of mature miR-198 in human trophoblasts. DISCUSSION: Our data indicate that placental FSTL1 is expressed particularly in extravillous trophoblasts. We also found no evidence for placental expression of miR-198, or for its regulation of FSTL1, implying that the post-transcriptional regulation of FSTL1 by miR-198 is tissue specific.
Authors: Masayuki Shimano; Noriyuki Ouchi; Kazuto Nakamura; Bram van Wijk; Koji Ohashi; Yasuhide Asaumi; Akiko Higuchi; David R Pimentel; Flora Sam; Toyoaki Murohara; Maurice J B van den Hoff; Kenneth Walsh Journal: Proc Natl Acad Sci U S A Date: 2011-10-10 Impact factor: 11.205
Authors: John T Hardy; Irina A Buhimschi; Megan E McCarthy; Guomao Zhao; Christine A Laky; Lydia L Shook; Catalin S Buhimschi Journal: J Clin Endocrinol Metab Date: 2016-05-09 Impact factor: 5.958