MicroRNA-1-3p inhibits the growth and metastasis of ovarian cancer cells by targeting DYNLT3

Eur Rev Med Pharmacol Sci. 2020 Sep;24(17):8713-8721. doi: 10.26355/eurrev_202009_22808.

Abstract

OBJECTIVE: The purpose of this study was to explore the role of microRNA-1-3p in the progression of ovarian cancer (OVC) and its possible molecular mechanisms.

Patients and methods: For the purpose of exploring the specific mechanism of the oncogene dynein light chain Tctex-type 3 (DYNLT3) in OVC, bioinformatics techniques were applied to predict miRNAs that might bind to DYNLT3, and then microRNA-1-3p was selected. After measuring the expression levels of microRNA-1-3p and DYNLT3 in 60 pairs of OVC tissue samples, the Pearson correlation analysis was used to calculate the expression correlation of microRNA-1-3p and DYNLT3. In addition, Dual-Luciferase reporting assay was used to verify the combination of the two in OVC cells. Furthermore, microRNA-1-3p NC, microRNA-1-3p mimics, and microRNA-1-3p mimics+ DYNLT3-OE (overexpression) were transfected into ES-2 and SKOV-3 cells, respectively. Subsequently, real-time quantitative polymerase chain reaction (qPCR) was performed to examine microRNA-1-3p level in each group of cells, followed by cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU) test, and transwell assay to verify the impact of microRNA-1-3p on the proliferation, migration ability, and invasiveness of OVC cells. Finally, the mRNA and protein levels of DYNLT3 were examined by qPCR and Western blot in OVC, respectively.

Results: Bioinformatics prediction results showed that a total of three possible miRNAs bound to the oncogene DYNLT3. Then, microRNA-1-3p was selected for further validation. qPCR results revealed that microRNA-1-3p was down-regulated in OVC tissues and cells, while DYNLT3 was up-regulated in OVC tissues. In addition, Pearson correlation analysis indicated that microRNA-1-3p was negatively correlated with DYNLT3 expression, and the Dual-Luciferase reporter assay confirmed that microRNA-1-3p was able to bind directly to the 3'-UTR of DYNLT3. Besides, microRNA-1-3p-mimics transfection remarkably decreased the mRNA and protein expressions of DYNLT3. On the contrary, transfection of microRNA-1-3p-mimics remarkably inhibited the proliferation, migration ability, and invasiveness of OVC cells. Moreover, the transfection of microRNA-1-3p-mimics+DYNLT3-OE partially reversed the inhibitory effect of microRNA-1-3p-mimics on the proliferative, migrate ability, and invasiveness of OVC cells.

Conclusions: MicroRNA-1-3p is under expressed either in OVC tissues or in cell lines, and overexpression of microRNA-1-3p may inhibit proliferative and migrate ability and invasiveness of OVC cells by modulating DYNLT3, which make microRNA-1-3p a potential therapeutic target for OVC.

MeSH terms

  • Cell Line
  • Cell Movement / genetics*
  • Cell Proliferation / genetics*
  • Dyneins / genetics*
  • Female
  • Humans
  • MicroRNAs / genetics*
  • Ovarian Neoplasms / genetics*
  • Ovarian Neoplasms / pathology*

Substances

  • MIRN1 microRNA, human
  • MicroRNAs
  • DYNLT3 protein, human
  • Dyneins