MicroRNA-330 inhibits growth and migration of melanoma A375 cells: In vitro study

J Cell Biochem. 2020 Jan;121(1):458-467. doi: 10.1002/jcb.29211. Epub 2019 Jun 25.

Abstract

Melanoma skin cancer is one of the main causes of male cancer-related deaths worldwide. It has been suggested that miR-330-5p can act as a tumor suppressor in various types of cancers. So, in this study, we replaced miR-330 in melanoma cancer cells by vector-based miR-330 to evaluate the effects of this microRNA on the growth and migration inhibition of melanoma cancer cells, and to determine the molecular mechanisms underlying its action. By using the MTT assay, the IC50 of Geneticin antibiotic was obtained as 460 µg/mL. The results of the qRT-PCR showed the increased expression level of miR-330 and decreased expression levels of MMP-9, CXCR4, Vimentin, melanoma cell adhesion molecule, AKT1, and E2F1 messenger RNA in A375 transfected cells. The cytotoxicity assay results demonstrated the inhibition of cancer cells proliferation. Furthermore, the wound healing test results showed a migration reduction of transfected cells with miR-330 compared with nontransfected ones. In addition, 4',6-diamidino-2-phenylindoleLB: Luria-Bertani (DAPI) staining revealed the significant nucleus fragmentation in miR-330 replaced cells, which correspond to apoptosis induction in replaced cells. The results showed that increase in miR-330 expression level could significantly inhibit the tumor cell growth and the migration of melanoma cancer cells.

Keywords: melanoma cancer; miR-330; migration; proliferation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism*
  • Cell Movement*
  • Cell Proliferation*
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • In Vitro Techniques
  • Matrix Metalloproteinase 9 / genetics
  • Matrix Metalloproteinase 9 / metabolism
  • Melanoma / genetics
  • Melanoma / metabolism
  • Melanoma / pathology*
  • MicroRNAs / genetics*
  • Receptors, CXCR4 / genetics
  • Receptors, CXCR4 / metabolism
  • Tumor Cells, Cultured
  • Vimentin / genetics
  • Vimentin / metabolism

Substances

  • Biomarkers, Tumor
  • CXCR4 protein, human
  • MIRN330 microRNA, human
  • MicroRNAs
  • Receptors, CXCR4
  • VIM protein, human
  • Vimentin
  • MMP9 protein, human
  • Matrix Metalloproteinase 9