Overexpressed miR-301a promotes cell proliferation and invasion by targeting RUNX3 in gastric cancer

J Gastroenterol. 2013 Sep;48(9):1023-33. doi: 10.1007/s00535-012-0733-6. Epub 2013 Jan 22.

Abstract

Background: MicroRNAs can promote or suppress the evolution of malignant behaviors by regulating multiple targets. We aimed to determine the expression of miR-301a recently screened in gastric cancer, to investigate the biological effects of miR-301a and to identify the specific miR-301a target gene.

Methods: Quantitative real-time RT-PCR was used to test miR-301a expression. Functional effects were explored by a water-soluble tetrazolium salt assay, a colony formation assay in soft agar, a migration assay, an invasion assay and cytometry used to determine apoptosis and cell cycle. Nude mice were inoculated subcutaneously by retrovirus-mediated stably expressed SGC-7901 cells. The target gene was determined by bioinformatic algorithms, dual luciferase reporter assay and Western blot.

Results: Firstly, we found that miR-301a was significantly upregulated both in cells and tissues of gastric cancer. The expression level of miR-301a was inversely correlated with tumor differentiation of gastric cancer tissues. Secondly, miR-301a promoted cell growth, soft agar clonogenicity, migration, invasion, and decreased cell apoptosis induced by cisplatin in vitro, while blockage of miR-301a reduced the percentage of G2/M phase cells via flow cytometry in gastric cancer cells. Ectopic expression of miR-301a enhanced the subcutaneous tumorigenesis in vivo. Finally, miR-301a directly downregulated RUNX3 expression post-transcriptionally in gastric cancer.

Conclusion: Our results demonstrate that miR-301a plays important roles in the development of gastric cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Cell Differentiation / physiology
  • Cell Line, Tumor
  • Cell Proliferation
  • Core Binding Factor Alpha 3 Subunit / biosynthesis*
  • Core Binding Factor Alpha 3 Subunit / genetics
  • Gene Expression Regulation, Neoplastic / physiology
  • Heterografts
  • Male
  • Mice
  • Mice, Inbred BALB C
  • MicroRNAs / biosynthesis*
  • MicroRNAs / genetics
  • Neoplasm Invasiveness
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplasm Staging
  • Neoplasm Transplantation
  • RNA, Neoplasm / biosynthesis
  • RNA, Neoplasm / genetics
  • Stomach Neoplasms / metabolism*
  • Stomach Neoplasms / pathology
  • Tumor Stem Cell Assay

Substances

  • Core Binding Factor Alpha 3 Subunit
  • MIRN301 microRNA, mouse
  • MicroRNAs
  • Neoplasm Proteins
  • RNA, Neoplasm