title: MicroRNA-340 Induces Apoptosis and Inhibits Metastasis of Ovarian Cancer Cells by Inactivation of NF-x03BA;B1.
abstract: OBJECTIVE
  Aberrant expression of microRNA-340 (miR-340) has been frequently reported in some cancers excluding ovarian cancer (OC). The role and its molecular mechanism of miR-340 in OC have not been reported.
  
  
  METHODS
  Real-time PCR was performed to detect the expression of miR-340 in OC cell lines. MiR-340 mimic and negative control were transfected into OC cells and the effects of miR-340 on the cell proliferation, cell cycle, apoptosis and metastasis were investigated by Brdu-ELISA assay, flow cytometry, qRT-PCR, Transwell and ELISA assays. Furthermore, protein level of NF-x03BA;B1 was measured by Western blotting. Meanwhile, luciferase assays were performed to validate NF-x03BA;B1 as miR-340 target in OC cells.
  
  
  RESULTS
  In this study, we explored the effects of miR-340 overexpression on apoptosis, invasion and EMT in OC cells. The mRNA level of miR-340 in OC cell lines and tissues was evidently reduced. The miR-340 mimic was transiently transfected into OC cells using Lipofectamine™ 2000 reagent. Subsequently, the Brdu-ELISA results showed that introduction of miR-340 inhibited cell proliferation. Our data also demonstrated that miR-340 mimic arrested cell cycle progression and promoted apoptosis of OC cells. In addition, miR-340 overexpression could also inhibit invasion and EMT of OC cells. qRT-PCR were used to determined the expressions of matrix metalloproteinase-2 and -9 (MMP-2 and -9) in OC cells. Next, we found that NF-x03BA;B1 expression was evidently reduced by up-regulation of miR-340. Bioinformatics analysis predicted that the NF-x03BA;B1 was a potential target gene of miR-340. Luciferase reporter assay further confirmed that miR-340 could directly target the 3' UTR of NF-x03BA;B1. Moreover, overexpression of NF-x03BA;B1 in OC cells transfected with miR-340 mimic partially reversed the inhibitory of miR-340 mimic.
  
  
  CONCLUSIONS
  miR-340 induced cell apoptosis and inhibited metastasis in OC cells by down-regulation of NF-x03BA;B1.
authors.forename: Peiquan
authors.initials: P
authors.lastname: Li
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Sun
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
mesh_terms: D020413:3' Untranslated Regions; D000070416:Antagomirs / Q000378:metabolism; D017209:Apoptosis; D001483:Base Sequence; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D015854:Up-Regulation
chemical_list: D020413:3' Untranslated Regions; D000070416:Antagomirs; D015820:Cadherins; D035683:MicroRNAs; D052002:NF-kappa B p50 Subunit; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9