Long noncoding RNASEH1-AS1 exacerbates the progression of non-small cell lung cancer by acting as a ceRNA to regulate microRNA-516a-5p/FOXK1 and thereby activating the Wnt/β-catenin signaling pathway

Cancer Med. 2022 Apr;11(7):1589-1604. doi: 10.1002/cam4.4509. Epub 2022 Feb 15.

Abstract

Background: Till now, no study has focused on the functions of RNASEH1 antisense RNA 1 (RNASEH1-AS1) in non-small cell lung cancer (NSCLC). Accordingly, we measured the expression of RNASEH1-AS1 in NSCLC and characterized its functions in detail. Finally, our research elucidated the mechanisms that occurred downstream of RNASEH1-AS1.

Methods: RNASEH1-AS1 expression was examined utilizing TCGA database and qRT-PCR. Functional experiments were conducted to study the tumor-associated functions of RNASEH1-AS1. The targeting relationship among RNASEH1-AS1, microRNA-516a-5p (miR-516a-5p), and forkhead box K1 (FOXK1) was revealed utilizing RNA immunoprecipitation and luciferase reporter assays.

Results: Utilizing TCGA database and our own cohort, we found a significantly increased level of RNASEH1-AS1 in NSCLC. The high level of RNASEH1-AS1 was markedly related with poor clinical outcomes. Knockdown of RNASEH1-AS1 expression inhibited NSCLC cell growth, metastatic capacities, and epithelial-mesenchymal transition and promoted the apoptosis in vitro, whereas RNASEH1-AS1 overexpression exerted the opposite effects. Additionally, knocking down RNASEH1-AS1 expression suppressed tumor growth in vivo. RNASEH1-AS1 was confirmed to act as a miR-516a-5p sponge, consequently upregulating FOXK1 expression in NSCLC cells. As revealed by the subsequent rescue experiments, the miR-516a-5p/FOXK1 axis served as a downstream effector of RNASEH1-AS1. In addition, by controlling the miR-516a-5p/FOXK1 axis, RNASEH1-AS1 was capable of activating the Wnt/β-catenin pathway.

Conclusion: RNASEH1-AS1 exacerbated the oncogenicity of NSCLC by affecting the miR-516a-5p/FOXK1 axis and consequently promoting the activation of Wnt/β-catenin pathway. Our newly identified RNASEH1-AS1/miR-516a-5p/FOXK1/Wnt/β-catenin network may offer an interesting foundation for NSCLC treatment in the clinic.

Keywords: RNASEH1 antisense RNA 1; Wnt/β-catenin pathway; ceRNA network; epithelial-mesenchymal transition.

MeSH terms

  • Carcinoma, Non-Small-Cell Lung* / genetics
  • Carcinoma, Non-Small-Cell Lung* / pathology
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • Forkhead Transcription Factors / genetics
  • Forkhead Transcription Factors / metabolism
  • Humans
  • Lung Neoplasms* / genetics
  • Lung Neoplasms* / pathology
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • RNA, Long Noncoding* / genetics
  • RNA, Long Noncoding* / metabolism
  • Wnt Signaling Pathway*
  • beta Catenin / genetics
  • beta Catenin / metabolism

Substances

  • CTNNB1 protein, human
  • FOXK1 protein, human
  • Forkhead Transcription Factors
  • MIRN516 microRNA, human
  • MicroRNAs
  • RNA, Long Noncoding
  • beta Catenin