Circ-10720 as a ceRNA adsorbs microRNA-1238 and modulates ZEB2 to boost NSCLC development by activating EMT

Eur J Med Res. 2024 Apr 12;29(1):226. doi: 10.1186/s40001-024-01715-9.

Abstract

Background: Circular RNAs (circRNAs) are critical regulators in the progression of tumors. This experimental design aimed to explore the mechanism of circ-10720 in non-small cell lung cancer (NSCLC).

Methods: We used RT-qPCR to measure circ-10720 expression in clinical samples and analyzed its relationship with the clinicopathological characteristics of NSCLC patients. The expression levels of microRNA-1238 (miR-1238) and Zinc Finger E-box-binding Homeobox 2 (ZEB2) in clinical samples were detected by RT-qPCR. NSCLC cells were transfected with relevant plasmids or sequences. Circ-10720, miR-1238, and ZEB2 expressions in cells were analyzed via RT-qPCR or western blot. Cell proliferation, apoptosis, migration, and invasion were assessed with CCK-8, flow cytometry, and transwell assay, respectively. The protein expression of ZEB2 and epithelial-mesenchymal transition (EMT)-related markers (E-cadherin, Vimentin, N-cadherin) were detected via western blot. Xenograft assay was used to determine the effect of circ-10720 on NSCLC in vivo. Circ-10720 and ZEB2 expressions in tumors were detected using RT-qPCR or Western blot. Immunohistochemistry was used to evaluate E-cadherin and N-cadherin expression in tumors. Finally, the binding relationship between miR-1238 with circ-10720 or ZEB2 was verified by the bioinformatics website, dual luciferase reporter assay, RNA pull-down assay, and RIP assay.

Results: Circ-10720 was upregulated in NSCLC and correlated with TNM stage of NSCLC patients. MiR-1238 was lowly expressed but ZEB2 was highly expressed in NSCLC. Circ-10720 silencing suppressed the proliferation, metastasis, and EMT of NSCLC cells. Mechanically, circ-10720 was a competitive endogenous RNA (ceRNA) for miR-1238, and ZEB2 was a target of miR-1238. circ-10720-modulated ZEB2 via competitively binding with miR-1238 to control NSCLC progression. In addition, circ-10720 knockdown suppressed tumor growth in vivo.

Conclusions: Circ-10720 acts as a ceRNA to adsorb miR-1238 and modulate ZEB2 to facilitate the proliferation, migration, invasion, and EMT of NSCLC cells.

Keywords: Circular RNA-10720; Epithelial–mesenchymal transition; MicroRNA-1238; Non-small cell lung cancer; Zinc Finger E-box-binding Homeobox 2.

MeSH terms

  • Cadherins
  • Carcinoma, Non-Small-Cell Lung* / genetics
  • Epithelial-Mesenchymal Transition / genetics
  • Humans
  • Lung Neoplasms* / genetics
  • MicroRNAs* / genetics
  • RNA, Circular
  • RNA, Competitive Endogenous
  • Zinc Finger E-box Binding Homeobox 2 / genetics

Substances

  • Cadherins
  • MicroRNAs
  • MIRN1238 microRNA, human
  • RNA, Competitive Endogenous
  • ZEB2 protein, human
  • Zinc Finger E-box Binding Homeobox 2
  • RNA, Circular