Document Detail


AKT isoforms 1 and 3 regulate basal and epidermal growth factor-stimulated SGHPL-5 trophoblast cell migration in humans.
MedLine Citation:
PMID:  23303682     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Protein kinase B/AKT is critically involved in murine placental development and migration of human placental trophoblasts into maternal uterine tissue. However, localization of the three AKT isoforms within human placenta and their roles in extravillous trophoblasts have not been elucidated. Therefore, we analyzed the expression pattern and function of AKT1, AKT2, and AKT3 in migratory human trophoblasts using SGHPL-5 cell pools stably expressing small-hairpin microRNA (shRNAmir) against AKT1, AKT2, or AKT3 as a model. Western blot analyses using isoform-specific antibodies revealed ubiquitous expression of AKT1, AKT2, and AKT3 in primary villous and extravillous trophoblasts and the trophoblastic cell lines JEG-3, HTR-8/SVneo, and SGHPL-5. Immunofluorescence of first-trimester placentae localized AKT2 and AKT3 to the cytoplasm and nucleus, respectively, in all subtypes of cytotrophoblasts, whereas AKT1 was detected in both cellular compartments. A similar distribution of AKT isoforms was detectable in SGHPL-5 cells. Gene silencing using shRNAmir decreased protein expression of AKT1, AKT2, and AKT3 to 16%, 8%, and 11%, respectively, in SGHPL-5 cells. Compared with shRNAmir controls, proliferation and camptothecin-induced apoptosis were not affected in the different AKT knockdown cells. However, basal and epidermal growth factor (EGF)-induced trophoblast migration was significantly reduced in AKT1 and AKT3 gene-silenced cells, whereas downregulation of AKT2 was not effective. Accordingly, a decrease in EGF-stimulated phosphorylation of AKT (Ser473 and Thr308) and its downstream target mTORC1 (Ser2448) was noticed in AKT1 and AKT3 shRNAmir cell pools. In summary, the results suggest that the AKT isoforms 1 and 3 promote basal as well as EGF-induced trophoblast migration.
Authors:
Peter Haslinger; Sandra Haider; Stefan Sonderegger; Jan Velten Otten; Jürgen Pollheimer; Guy Whitley; Martin Knöfler
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2013-03-07
Journal Detail:
Title:  Biology of reproduction     Volume:  88     ISSN:  1529-7268     ISO Abbreviation:  Biol. Reprod.     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-03-08     Completed Date:  2013-09-17     Revised Date:  2013-09-30    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  United States    
Other Details:
Languages:  eng     Pagination:  54     Citation Subset:  IM    
Affiliation:
Department of Obstetrics and Fetal-Maternal Medicine, Reproductive Biology Unit, Medical University of Vienna, Vienna, Austria.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis
Camptothecin
Cell Line, Tumor
Cell Movement*
Cell Proliferation
Down-Regulation
Epidermal Growth Factor / metabolism
Female
Gene Expression
Gene Silencing
Humans
MicroRNAs
Phosphorylation
Pregnancy
Protein Isoforms / metabolism
Proto-Oncogene Proteins c-akt / metabolism*
TOR Serine-Threonine Kinases / metabolism
Topoisomerase I Inhibitors
Trophoblasts / metabolism*
Chemical
Reg. No./Substance:
0/MicroRNAs; 0/Protein Isoforms; 0/Topoisomerase I Inhibitors; 0/mechanistic target of rapamycin complex 1; 62229-50-9/Epidermal Growth Factor; 7689-03-4/Camptothecin; EC 2.7.1.-/AKT3 protein, human; EC 2.7.1.1/TOR Serine-Threonine Kinases; EC 2.7.11.1/AKT1 protein, human; EC 2.7.11.1/AKT2 protein, human; EC 2.7.11.1/Proto-Oncogene Proteins c-akt

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