LncRNA TP73-AS1 regulates miR-495 expression to promote migration and invasion of nasopharyngeal carcinoma cells through junctional adhesion molecule A

Kaohsiung J Med Sci. 2021 May;37(5):361-370. doi: 10.1002/kjm2.12338. Epub 2021 Jan 5.

Abstract

The main obstacle to the treatment of nasopharyngeal carcinoma (NPC) is metastasis. Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) are highly involved in the progression of NPC. In this study, we aimed to explore the regulatory role of lncRNA P73 antisense RNA 1 T (TP73-AS1) and miR-495 in migration and invasion of NPC cells. The expression levels of TP73-AS1, miR-495, and junctional adhesion molecule A (JAM-A) in NPC tissue samples and cell lines were examined by quantitative real-time PCR (qRT-PCR) and/or Western blot. NPC cells were transfected with vectors overexpressing TP73-AS1, short hairpin RNA (shRNA) against TP73-AS1, shRNA against JAM-A, miR-495 mimics, miR-495 inhibitor, and their corresponding negative controls as designated. The MTT assay, cell migration assay, and transwell assay were performed to detect cell viability, migration, and invasion, respectively. Dual-luciferase reporter assay was performed to confirm the binding of TP73-AS1 and miR-495, and miR-495 and JAM-A. TP73-AS1 and JAM-A were significantly upregulated while miR-495 was markedly downregulated in NPC tissues and cell lines compared to normal controls. The overexpression of TP73-AS1 promoted migration and invasion of NPC cell line CNE-2. TP73-AS1 targeted miR-495 and negatively regulated its expression. TP73-AS1 upregulated the expression of JAM-A through miR-495. TP73-AS1 mediated migration and invasion of CNE-2 cells via upregulating JAM-A. LncRNA TP73-AS1, miR-495, and JAM-A are involved in migration and invasion of NPC cells. The TP73-AS1/miR-495/JAM-A axis may serve as a therapeutic target for the treatment of NPC.

Keywords: invasion; long non-coding RNA TP73-AS1; microRNA-495; migration; nasopharyngeal carcinoma.

MeSH terms

  • Biopsy
  • Cell Adhesion Molecules / metabolism*
  • Cell Line, Tumor
  • Cell Movement
  • Disease Progression
  • Epithelial-Mesenchymal Transition
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • MicroRNAs / biosynthesis*
  • Nasopharyngeal Carcinoma / metabolism*
  • Nasopharyngeal Neoplasms / metabolism*
  • Neoplasm Invasiveness
  • Neoplasm Metastasis
  • Protein Binding
  • RNA, Long Noncoding / physiology*
  • Receptors, Cell Surface / metabolism*
  • Tetrazolium Salts / chemistry
  • Thiazoles / chemistry

Substances

  • Cell Adhesion Molecules
  • F11R protein, human
  • MIRN495 microRNA, human
  • MicroRNAs
  • RNA, Long Noncoding
  • Receptors, Cell Surface
  • Tetrazolium Salts
  • Thiazoles
  • long non-coding RNA KIAA0495, human
  • thiazolyl blue