title: Prostaglandin E2-mediated migration of human trophoblast requires RAC1 and CDC42.
abstract: The invasion of maternal decidua and uterine spiral arteries by a trophoblast subpopulation called extravillous trophoblast (EVT) is essential for the establishment of a normal placenta and an adequate blood flow toward the fetus. Derangements in these processes underlie pregnancy-related diseases like preeclampsia and intrauterine growth restriction. Many growth factors, growth factor binding proteins, and extracellular matrix components can positively or negatively regulate the proliferation, migration, and/or invasiveness of these EVT cells. RHO GTPases, including RHOA, RAC1, and CDC42, are ubiquitous proteins that control cytoskeletal changes by forming stress fibers and projecting lamellipodia and filopodia during cellular migration. We had previously shown that prostaglandin (PG) E(2) produced in abundance by the decidua promotes the migration of first-trimester human EVTs by increasing the intracellular concentration of calcium and activating calpain. Using our well-characterized immortalized EVT cell line, HTR-8/SVneo, as well as villus explants from first-trimester placentae, this study examined the role of RHO GTPases RAC1 and CDC42 in PGE(2)-mediated migratory responses of these cells. Though a RAC1 inhibitor, NSC23766 as well as RAC1 knockdown by siRNA decreased the migration of HTR-8/SVneo cells in a Transwell migration assay, this inhibition could not be restored by PGE(2) or 17-phenyl trinor PGE(2) (PGE receptor PTGER1 agonist) or PGE(1) Alcohol (PGE receptor PTGER4 agonist). Similar results were noted for EVT cell spreading in villus explants. Furthermore, CDC42 silencing using siRNA inhibited PGE(2)-induced migration of HTR-8/SVneo cells. Finally, the treatment of EVT cells with PGE(2), PTGER1 agonist, or PTGER4 agonist activated RAC1 and CDC42 at 10 min, suggesting that RAC1 and CDC42 play an essential role in PGE(2)-mediated migration of human EVTs.
authors.affiliation: Departments of Anatomy and Cell Biology and Pathology, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada N6A 5C1.
authors.forename: Catalin
authors.initials: C
authors.lastname: Nicola
authors.forename: Peeyush K
authors.initials: PK
authors.lastname: Lala
authors.forename: Chandan
authors.initials: C
authors.lastname: Chakraborty
mesh_terms: D000634:Aminoquinolines / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D015232:Dinoprostone / Q000494:pharmacology / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D011247:Pregnancy; D011743:Pyrimidines / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D046509:Tissue Culture Techniques; D014327:Trophoblasts / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D020764:cdc42 GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D020830:rac1 GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*
chemical_list: D000634:Aminoquinolines; D004791:Enzyme Inhibitors; C487513:NSC 23766; D011743:Pyrimidines; C107953:RAC1 protein, human; D034741:RNA, Small Interfering; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D015232:Dinoprostone