-19-0088 159 Silencing SEC5 inhibits trophoblast invasion via integrin/Ca 2+ signaling

The invasion of maternal decidua by extravillous trophoblast (EVT) is essential for the establishment and maintenance of pregnancy, and abnormal trophoblast invasion could lead to placenta-associated pathologies including early pregnancy loss and preeclampsia. SEC5, a component of the exocyst complex, plays important roles in cell survival and migration, but its role in early pregnancy has not been reported. Thus, the present study was performed to explore the functions of SEC5 in trophoblast cells. The results showed that SEC5 expression in human placental villi at first trimester was significantly higher than it was at the third trimester, and it was abundantly localized in the cytotrophoblast (CTB) and the trophoblastic column. SEC5 knockdown was accompanied by reduced migration and invasion in HTR-8/SVneo cells. In addition, the expression and plasma membrane distribution of integrin β 1 was also decreased. Furthermore, shRNA-mediated knockdown of SEC5 inhibited the outgrowth of first trimester placental explants. SEC5 and InsP 3 R were colocalized in the cytoplasm of HTR-8/SVneo cells, and the cell-permeant calcium chelator BAPTA-AM could significantly inhibit HTR-8/SVneo cell invasion. The Ca 2+ imaging results showed that the 10% fetal bovine serum-stimulated cytosolic calcium concentration ([Ca 2+ ]c) was not only reduced by downregulated SEC5 but also was blocked by the InsP 3 R inhibitor. Furthermore, either the [Ca 2+ ]c was buffered by BAPTA-AM or the knockdown of SEC5 disrupted HTR-8/SVneo cell F-actin stress fibers and caused cytoskeleton derangement. Taken together, our results suggest that SEC5 might be involved in regulating trophoblast cell migration and invasion through the integrin/Ca 2+ signal pathway to induce cytoskeletal rearrangement.

[1]  Haiyang Yu,et al.  Cell Biology International , 2019 .

[2]  J. Pollheimer,et al.  Regulation of Placental Extravillous Trophoblasts by the Maternal Uterine Environment , 2018, Front. Immunol..

[3]  Hui-qin Mo,et al.  EIF5A1 promotes trophoblast migration and invasion via ARAF-mediated activation of the integrin/ERK signaling pathway , 2018, Cell Death & Disease.

[4]  Yan Gu,et al.  miR-3074-5p Promotes the Apoptosis but Inhibits the Invasiveness of Human Extravillous Trophoblast–Derived HTR8/SVneo Cells In Vitro , 2018, Reproductive Sciences.

[5]  Lan Yan,et al.  InsP3R-SEC5 interaction on phagosomes modulates innate immunity to Candida albicans by promoting cytosolic Ca2+ elevation and TBK1 activity , 2018, BMC Biology.

[6]  Lan Yan,et al.  InsP 3 R-SEC 5 interaction on phagosomes modulates innate immunity to Candida albicans by promoting cytosolic Ca 2 + elevation and TBK 1 activity , 2018 .

[7]  Ching-Wen Chang,et al.  Trophoblast lineage specification, differentiation and their regulation by oxygen tension. , 2018, The Journal of endocrinology.

[8]  Toshiaki Tanaka,et al.  Diverse Functions and Signal Transduction of the Exocyst Complex in Tumor Cells , 2017, Journal of cellular physiology.

[9]  J. Aplin,et al.  Embryo–epithelium interactions during implantation at a glance , 2017, Journal of Cell Science.

[10]  P. Schneider,et al.  Integrin-mediated transactivation of P2X7R via hemichannel-dependent ATP release stimulates astrocyte migration. , 2016, Biochimica et biophysica acta.

[11]  Hee-Jung Choi,et al.  Integrin αVβ3 and αVβ5 are required for leukemia inhibitory factor-mediated the adhesion of trophoblast cells to the endometrial cells. , 2016, Biochemical and biophysical research communications.

[12]  O. Lee,et al.  Dexamethasone-induced cellular tension requires a SGK1-stimulated Sec5–GEF-H1 interaction , 2015, Journal of Cell Science.

[13]  Yan Gu,et al.  Aberrant Placental Villus Expression of miR-486-3p and miR-3074-5p in Recurrent Miscarriage Patients and Uterine Expression of These MicroRNAs during Early Pregnancy in Mice , 2015, Gynecologic and Obstetric Investigation.

[14]  D. D. Mak,et al.  Inositol 1,4,5-trisphosphate receptors in the endoplasmic reticulum: A single-channel point of view. , 2015, Cell calcium.

[15]  Hellyeh Hamidi,et al.  Integrin traffic – the update , 2015, Journal of Cell Science.

[16]  Xiaojun Tan,et al.  A Kinase-Independent Role for EGF Receptor in Autophagy Initiation , 2015, Cell.

[17]  J. Robinson,et al.  Exocyst complex protein expression in the human placenta. , 2014, Placenta.

[18]  Michael L. Gatza,et al.  TβRIII/β-arrestin2 regulates integrin α5β1 trafficking, function, and localization in epithelial cells , 2013, Oncogene.

[19]  M. White,et al.  The microtubule-associated Rho activating factor GEF-H1 interacts with exocyst complex to regulate vesicle traffic. , 2012, Developmental cell.

[20]  J. Eun,et al.  Dynamic alterations in integrin α4 expression by hypoxia are involved in trophoblast invasion during early implantation , 2012, Journal of cellular biochemistry.

[21]  Suyong Choi,et al.  Phosphoinositide signaling regulates the exocyst complex and polarized integrin trafficking in directionally migrating cells. , 2012, Developmental cell.

[22]  H. Taylor,et al.  Implantation failure: molecular mechanisms and clinical treatment. , 2011, Human reproduction update.

[23]  K. Lim,et al.  Sec5 and Exo84 Foster Oncogenic Ras-Mediated Tumorigenesis , 2010, Molecular Cancer Research.

[24]  Y. Marikawa,et al.  Establishment of trophectoderm and inner cell mass lineages in the mouse embryo , 2009, Molecular reproduction and development.

[25]  E. Lengyel,et al.  Up-regulation of alpha5-integrin by E-cadherin loss in hypoxia and its key role in the migration of extravillous trophoblast cells during early implantation. , 2009, Endocrinology.

[26]  J. Kingdom,et al.  Glial cell missing-1 transcription factor is required for the differentiation of the human trophoblast , 2009, Cell Death and Differentiation.

[27]  C. Yeaman,et al.  Ral-regulated interaction between Sec5 and paxillin targets Exocyst to focal complexes during cell migration , 2008, Journal of Cell Science.

[28]  C. Crum,et al.  Pathophysiology of placenta creta: the role of decidua and extravillous trophoblast. , 2008, Placenta.

[29]  J. Cross,et al.  Diverse subtypes and developmental origins of trophoblast giant cells in the mouse placenta. , 2007, Developmental biology.

[30]  E. Staun-ram,et al.  Reproductive Biology and Endocrinology Open Access Human Trophoblast Function during the Implantation Process , 2022 .

[31]  G. Giannone,et al.  Calcium Oscillations Trigger Focal Adhesion Disassembly in Human U87 Astrocytoma Cells* , 2002, The Journal of Biological Chemistry.

[32]  P. Greer,et al.  Reduced Cell Migration and Disruption of the Actin Cytoskeleton in Calpain-deficient Embryonic Fibroblasts* , 2001, The Journal of Biological Chemistry.

[33]  A. Tarnawski,et al.  Nonsteroidal anti-inflammatory drugs inhibit re-epithelialization of wounded gastric monolayers by interfering with actin, Src, FAK, and tensin signaling. , 2001, Life sciences.

[34]  M. Brown,et al.  Paxillin-ARF GAP signaling and the cytoskeleton. , 2001, Current opinion in cell biology.

[35]  J. Kingdom,et al.  Macrophage-Induced Apoptosis Limits Endovascular Trophoblast Invasion in the Uterine Wall of Preeclamptic Women , 2001, Laboratory Investigation.

[36]  P. Lala,et al.  Insulin-like growth factor-binding protein 1 stimulates human trophoblast migration by signaling through alpha 5 beta 1 integrin via mitogen-activated protein Kinase pathway. , 2001, The Journal of clinical endocrinology and metabolism.

[37]  A. Campana,et al.  Paracrine and autocrine regulators of trophoblast invasion--a review. , 2000, Placenta.

[38]  Sean P. Palecek,et al.  Regulation of Cell Migration by the Calcium-dependent Protease Calpain* , 1997, The Journal of Biological Chemistry.

[39]  Daniel Choquet,et al.  Ligand binding regulates the directed movement of β1 integrins on fibroblasts , 1996, Nature.

[40]  J. Irving,et al.  Functional role of cell surface integrins on human trophoblast cell migration: regulation by TGF-beta, IGF-II, and IGFBP-1. , 1995, Experimental cell research.

[41]  K. Lim,et al.  Integrin switching regulates normal trophoblast invasion. , 1994, Development.

[42]  J. Spudich,et al.  Nonmuscle contractile proteins: the role of actin and myosin in cell motility and shape determination. , 1977, Annual review of biochemistry.