miR-342-3p affects hepatocellular carcinoma cell proliferation via regulating NF-κB pathway

Biochem Biophys Res Commun. 2015 Feb 13;457(3):370-7. doi: 10.1016/j.bbrc.2014.12.119. Epub 2015 Jan 9.

Abstract

Recent research indicates that non-coding microRNAs (miRNAs) help regulate basic cellular processes in many types of cancer cells. We hypothesized that overexpression of miR-342-3p might affect proliferation of hepatocellular carcinoma (HCC) cells. After confirming overexpression of miR-342-3p with qRT-PCR, MTT assay showed that HCC cell proliferation was significantly inhibited by miR-342-3p, and that it significantly decreased BrdU-positive cell proliferation by nearly sixfold. Searching for targets using three algorithms we found that miR-342-3p is related to the NF-κB pathway and luciferase assay found that IKK-γ, TAB2 and TAB3 are miR-342-3p target genes. Results of western blot on extracted nuclear proteins of HepG2 and HCT-116 cells showed that miR-342-3p reduced and miR-342-3p-in increased p65 nuclear levels and qRT-PCR found that NF-κB pathway downstream genes were downregulated by miR-342-3p and upregulated by miR-342-3p-in, confirming that miR-342 targets NF-κB pathway. Overexpression of Ikk-γ, TAB2 and TAB3 partially rescued HCC cells proliferation inhibited by miR-342-3p. Using the GSE54751 database we evaluated expression from 10 HCC samples, which strongly suggested downregulation of miR-342-3p and we also found inverse expression between miR-342-3p and its targets IKK-γ, TAB2 and TAB3 from 71 HCC samples. Our results show that miR-342-3p has a significant role in HCC cell proliferation and is suitable for investigation of therapeutic targets.

Keywords: Cell proliferation; Hepatocellular carcinoma; NF-κB signaling; miR-342-3p.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Base Sequence
  • Carcinoma, Hepatocellular / genetics*
  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology*
  • Cell Proliferation
  • Gene Expression Regulation, Neoplastic
  • HCT116 Cells
  • Hep G2 Cells
  • Humans
  • I-kappa B Kinase / genetics
  • Intracellular Signaling Peptides and Proteins / genetics
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology*
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism*
  • NF-kappa B / metabolism*
  • Signal Transduction
  • Transcription Factor RelA / metabolism
  • Transfection

Substances

  • Adaptor Proteins, Signal Transducing
  • Intracellular Signaling Peptides and Proteins
  • MIRN342 microRNA, human
  • MicroRNAs
  • NF-kappa B
  • RELA protein, human
  • TAB2 protein, human
  • TAB3 protein, human
  • Transcription Factor RelA
  • I-kappa B Kinase