Effects of CDKN2B-AS1 on cellular proliferation, invasion and AKT3 expression are attenuated by miR-424-5p in a model of ovarian endometriosis

Reprod Biomed Online. 2021 Jun;42(6):1057-1066. doi: 10.1016/j.rbmo.2021.02.004. Epub 2021 Feb 12.

Abstract

Research question: Endometriosis is a common and complicated gynaecologic disease. Long non-coding RNA CDKN2B-AS1 plays a crucial role in the development and progression of several cancers. Whether CDKN2B-AS1 contributes to endometriosis, however, remains unknown.

Design: Cellular proliferation, invasion and DNA synthesis abilities were assessed by CCK8, transwell and 5-ethynyle-2'-deoxyuridine assays. The expression of epithelial-mesenchymal transition markers and three isoforms of AKT was detected using Western blot. Real-time polymerase chain reaction was used to determine the relative expression levels of CDKN2B-AS1 and candidate miRNAs in ectopic, eutopic endometria and normal endometrial tissues. The relationship between CDKN2B-AS1 and miRNA was determined by luciferase reporter assays.

Results: The relative expression level of CDKN2B-AS1 was up-regulated in eutopic and ectopic endometria. In endometrial stromal cells and Ishikawa cells, CDKN2B-AS1 overexpression promoted cellular proliferation and invasion, and increased the protein expression of vimentin but decreased the expression of E-cadherin. miR-424-5p was confirmed the target of CDKN2B-AS1 through bioinformatics tools and luciferase reporter assays. In addition, the enhanced effect of cellular phenotype of CDKN2B-AS1 overexpression was significantly attenuated by miR-424-5p overexpression. Furthermore, miR-424-5p was able to directly target AKT3 through luciferase reporter assay. Mechanistically, CDKN2B-AS1 acts as a ceRNA by sponging miR-424-5p and targets AKT3.

Conclusions: The cellular mechanism of CDKN2B-AS1 in endometriosis was confirmed; CDKN2B-AS1 may be a potential target for ovarian endometriosis therapy.

Keywords: AKT3; CDKN2B-AS1; Epithelial–mesenchymal transition (EMT); Ovarian endometriosis; miR-424-5p.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Endometriosis / etiology
  • Endometriosis / metabolism*
  • Epithelial-Mesenchymal Transition
  • Female
  • Humans
  • MicroRNAs / metabolism*
  • Middle Aged
  • Ovarian Diseases / etiology
  • Ovarian Diseases / metabolism*
  • Primary Cell Culture
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RNA, Long Noncoding / metabolism*
  • Young Adult

Substances

  • CDKN2B antisense RNA, human
  • MIRN424 microrna, human
  • MicroRNAs
  • RNA, Long Noncoding
  • AKT3 protein, human
  • Proto-Oncogene Proteins c-akt