title: RNA interference-mediated knockdown of Notch-1 inhibits migration and invasion, down-regulates matrix metalloproteinases and suppresses NF-κB signaling pathway in trophoblast cells.
abstract: Preeclampsia is well known to present with reduced trophoblast invasion into the placental bed. Notch-1, a ligand-activated transmembrane receptor, has been reported to be down-regulated in preeclamptic human placentas. This study was conducted to explore the role of Notch-1 in the cell migration and invasion of a human trophoblast cell line, JEG3 cells. Short hairpin RNA (shRNA)-mediated RNA interference was performed to effectively suppress the endogenous expression of Notch-1 at both mRNA and protein levels in JEG3 cells. Results of wound healing and transwell assays showed that knockdown of Notch-1 reduced trophoblast cell migration and invasion. The protein expressions and activities of matrix metalloproteinase (MMP)-2 and MMP-9 were reduced in JEG3 cell when Notch-1 was decreased. Furthermore, the epithelial-cadherin (E-cadherin) expression increased in JEG3 cells when Notch-1 was inhibited, whereas its suppressor Snail decreased in these cells. Moreover, knockdown of Notch-1 also suppressed NF-κB signaling pathway by inhibiting the phosphorylation of nuclear factor kappa B (NF-κB p65) inhibitor (IκBα) and the subsequent nuclear translocation of NF-κB subunit p65 in JEG3 cells. In summary, we demonstrate that Notch-1 contributes to trophoblast cell migration and invasion and that it may be involved in the pathology of human preeclampsia.
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, People's Republic of China.
authors.forename: Leilei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Gynecology, Liaoning Cancer Hospital and Institute, Shenyang 110042, People's Republic of China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Tang
authors.affiliation: Department of Obstetrics and Gynecology, The Women and Infants Hospital of Shenyang City, Shenyang 110014, People's Republic of China.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, People's Republic of China. Electronic address: cmu2hshangtao@163.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Shang
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D015536:Down-Regulation / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D055785:Gene Knockdown Techniques; D006801:Humans; D020782:Matrix Metalloproteinases / Q000235:genetics* / Q000378:metabolism*; D009361:Neoplasm Invasiveness / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000037:antagonists & inhibitors / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics; D014327:Trophoblasts / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D000097782:NF-kappaB-Inducing Kinase
chemical_list: D020782:Matrix Metalloproteinases; D017346:Protein Serine-Threonine Kinases; D051881:Receptor, Notch1; D000097782:NF-kappaB-Inducing Kinase
keywords: Cell invasion; Cell migration; Notch-1; Preeclampsia; Trophoblast cells; shRNA