Up-regulation of miR-10b-3p promotes the progression of hepatocellular carcinoma cells via targeting CMTM5

J Cell Mol Med. 2018 Jul;22(7):3434-3441. doi: 10.1111/jcmm.13620. Epub 2018 Apr 24.

Abstract

In this study, we investigated how miR-10b-3p regulated the proliferation, migration, invasion in hepatocellular carcinoma (HCC) at both in vitro and in vivo levels. CMTM5 was among the differentially expressed genes (data from TCGA). The expression of miR-10b-3p and CMTM5 was detected by qRT-PCR and Western blot (WB). TargetScan was used to acquire the binding sites. Dual-luciferase reporter gene assay was used to verify the direct target relationship between miR-10b-3p and CMTM5. WB analysis proved that miR-10b-3p suppressed CMTM5 expression. Furthermore, proliferation, invasion and migration of HCC cells were measured by MTT assay, colony formation assay, transwell assay and wound-healing assay, respectively. Kaplan-Meier plotter valued the overall survival of CMTM5. Finally, xenograft assay was also conducted to verify the effects of miR-10b-3p/CMTM5 axis in vivo. Up-regulation of miR-10b-3p and down-regulation of CMTM5 were detected in HCC tissues and cell lines. CMTM5 was verified as a target gene of miR-10b-3p. The overexpression of CMTM5 contributed to the suppression of the proliferative, migratory and invasive abilities of HCC cells. Moreover, the up-regulation of miR-10b-3p and down-regulation of CMTM5 were observed to be associated with worse overall survival. Lastly, we have confirmed the carcinogenesis-related roles of miR-10b-3p and CMTM5 in vivo. We concluded that the up-regulation of miR-10b-3p promoted the progression of HCC cells via targeting CMTM5.

Keywords: CMTM5; hepatocellular carcinoma; miR-10b-3p.

MeSH terms

  • Animals
  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / mortality
  • Carcinoma, Hepatocellular / pathology*
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Chemokines / genetics*
  • Chemokines / metabolism
  • Female
  • Gene Expression Regulation, Neoplastic
  • Hep G2 Cells
  • Humans
  • Liver Neoplasms / genetics
  • Liver Neoplasms / mortality
  • Liver Neoplasms / pathology*
  • MARVEL Domain-Containing Proteins / genetics*
  • MARVEL Domain-Containing Proteins / metabolism
  • Mice, Inbred BALB C
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Survival Analysis
  • Tumor Suppressor Proteins / genetics*
  • Tumor Suppressor Proteins / metabolism

Substances

  • CMTM5 protein, human
  • Chemokines
  • MARVEL Domain-Containing Proteins
  • MIRN10 microRNA, human
  • MicroRNAs
  • Tumor Suppressor Proteins