title: NF-κB-modulated miR-130a targets TNF-α in cervical cancer cells.
abstract: BACKGROUND
  Nuclear factor-κB (NF-κB) induces a variety of biological processes through transcriptional gene control whose products are components in various signaling pathways. MicroRNAs are a small endogenous non-coding RNAs that regulate gene expression and are involved in tumorigenesis. Using human cervical cancer cell lines, this study aimed to investigate whether NF-κB could regulate miR-130a expression and the functions and targets of miR-130a.
  
  
  METHODS
  We used the HeLa and C33A cervical cancer cell lines that were transfected with NF-κB or miR-130a overexpression plasmids to evaluate their effects on cell growth. We utilized bioinformatics, a fluorescent reporter assay, qRT-PCR and Western blotting to identify downstream target genes.
  
  
  RESULTS
  In HeLa and C33A cells, NF-κB and miR-130a overexpression promoted cell growth, but genetic knockdowns suppressed growth. TNF-α was identified as a target of miR-130a by binding in a 3'-untranslated region (3'UTR) EGFP reporter assay and by Western blot analysis. Furthermore, low TNF-α concentrations stimulated NF-κB activity and then induced miR-130a expression, and TNF-α overexpression rescued the effects of miR-130a on cervical cancer cells.
  
  
  CONCLUSIONS
  Our findings indicate that TNF-α can activate NF-κB activity, which can reduce miR-130a expression, and that miR-130a targets and downregulates TNF-α expression. Hence, we shed light on the negative feedback regulation of NF-κB/miR-130a/TNF-α/NF-κB in cervical cancer and may provide insight into the carcinogenesis of cervical cancer.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.forename: Haidong
authors.initials: H
authors.lastname: Wu
authors.forename: Pu
authors.initials: P
authors.lastname: Li
authors.forename: Yanzheng
authors.initials: Y
authors.lastname: Zhao
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.affiliation: Tianjin Life Science Research Center and School of Basic Medical Sciences, Tianjin Medical University, No, 22 Qi-Xiang-Tai Road, Tianjin 300070, China. htang2002@yahoo.com.
authors.forename: Hua
authors.initials: H
authors.lastname: Tang
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D005260:Female; D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D016328:NF-kappa B / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000378:metabolism* / Q000473:pathology
chemical_list: D017931:DNA Primers; C554019:MIRN130 microRNA, human; D035683:MicroRNAs; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha