Preliminary study on the effect of nucleolin specific aptamer-miRNA let-7d chimera on Janus kinase-2 expression level and activity in gastric cancer (MKN-45) cells

Mol Biol Rep. 2019 Feb;46(1):207-215. doi: 10.1007/s11033-018-4462-7. Epub 2018 Nov 10.

Abstract

Recently, much attention has been focused on the use of miRNAs in cancer treatment. The role of proto-oncogene Janus kinase-2 (JAK-2) in proliferation and survival of gastric cancer has been previously documented. The aim of this study was to evaluate the effect of a chimera consisted of nucleolin specific aptamer (NCL-Apt) and miRNA let-7d on JAK2 expression level and activity in gastric cancer cells. NCL-Apt-miRNA let-7d chimera was prepared by two methods. Gastric cancer (MKN-45) cell line and control cell line of human dermal fibroblast (HDF) were treated with the chimera and the changes in JAK2 expression and activity were determined using real-time PCR and ELISA techniques, respectively. In MKN-45 cells, the chimera caused significant decrease in JAK2 expression level and activity compared to the aptamer alone and miRNA mimic negative control. Nevertheless, transfected miRNA let-7d showed remarkable reduction in the expression level of JAK2 in comparison with control state in both MKN-45 and HDF, confirmed unspecific effect of let-7d on normal and cancerous cells. With regard to the synergic effect of this chimera on JAK2 activity, it might be viewed as a therapeutic candidate in gastric cancer. However, further studies are warranted to prove it.

Keywords: Gastric cancer; Janus kinase-2 (JAK-2); MiRNA let-7d; Nucleolin specific aptamer (NCL-Apt).

MeSH terms

  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / pharmacology
  • Cell Line, Tumor
  • Gene Expression Regulation / genetics
  • Gene Expression Regulation, Neoplastic / genetics
  • Humans
  • Janus Kinase 2 / genetics
  • Janus Kinase 2 / physiology
  • MicroRNAs* / genetics
  • MicroRNAs* / physiology
  • Nucleolin
  • Phosphoproteins* / genetics
  • Phosphoproteins* / physiology
  • Preliminary Data
  • Proto-Oncogene Mas
  • RNA-Binding Proteins* / genetics
  • RNA-Binding Proteins* / physiology
  • Stomach Neoplasms* / genetics

Substances

  • Aptamers, Nucleotide
  • JAK2 protein, human
  • Janus Kinase 2
  • MAS1 protein, human
  • MicroRNAs
  • mirnlet7 microRNA, human
  • Phosphoproteins
  • Proto-Oncogene Mas
  • RNA-Binding Proteins