title: [miR-497 suppresses proliferation of human cervical carcinoma HeLa cells by targeting cyclin E1].
abstract: OBJECTIVE
  To evaluate the effect of miR-497 on proliferation of human cervical carcinoma HeLa cells and target relationship between miR-497 and cyclin E1 (CCNE1).
  
  
  METHODS
  Pre-miR-497 sequences were synthesized and cloned into pcDNATM6.2-GW to construct recombinant plasmid pcDNATM6.2-GW-pre-miR-497 and identified by real-time quantitative PCR (qRT-PCR). In addition, sequences of the wild-type CCNE1 (WT-CCNE1) and mutant CCNE1 (MT-CCNE1) were respectively cloned into pmirGLO vectors. MTT assay was used to explore the impact of miR-497 on the proliferation of HeLa cells. Furthermore, the target effect of miR-497 on the CCNE1 was identified by dual-luciferase reporter assay system, qRT-PCR and Western blotting.
  
  
  RESULTS
  The recombinant plasmids pcDNATM6.2-GW-pre-miR-497 and pmirGLO-WT-CCNE1, pmirGLO-MT-CCNE1 were successfully constructed, and the miR-497 expression level in HeLa cells transfected with pre-miR-497 was significantly higher than that in the neg-miR group (P<0.05). MTT assay showed that miR-497 could significantly inhibit the proliferation of HeLa cells (P<0.05). A remarkable reduction of luciferase activities of WT-CCNE1 reporter was observed in HeLa cells with pre-miR-497 transfection (P<0.01), and the mRNA and protein expression levels of CCNE1 were down-regulated in HeLa cells transfected with pre-miR-497 (P<0.05).
  
  
  CONCLUSIONS
  Over-expressed miR-497 in HeLa cells could suppress cell proliferation by targeting CCNE1.
authors.affiliation: Medical College, Yan'an University, Yan'an 716000, China.
authors.forename: Jiming
authors.initials: J
authors.lastname: Han
authors.affiliation: Medical College, Yan'an University, Yan'an 716000, China.
authors.forename: Manpeng
authors.initials: M
authors.lastname: Huo
authors.affiliation: Medical College, Yan'an University, Yan'an 716000, China.
authors.forename: Mingtao
authors.initials: M
authors.lastname: Mu
authors.affiliation: Medical College, Yan'an University, Yan'an 716000, China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Medical College, Yan'an University, Yan'an 716000, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics; D019927:Cyclin E / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: C475820:CCNE1 protein, human; D019927:Cyclin E; C559640:MIRN497 microRNA, human; D035683:MicroRNAs; D015513:Oncogene Proteins; D008156:Luciferases