Circ_0014235 confers Gefitinib resistance and malignant behaviors in non-small cell lung cancer resistant to Gefitinib by governing the miR-146b-5p/YAP/PD-L1 pathway

Cell Cycle. 2022 Jan;21(1):86-100. doi: 10.1080/15384101.2021.2009986. Epub 2021 Dec 17.

Abstract

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), such as Gefitinib, have been recommended as the first-line treatment reagent for advanced EGFR-mutant non-small cell lung cancer (NSCLC). However, the mechanisms of drug resistance development are not fully determined. This study aimed to explore the role of circular RNA (circ_0014235) in Gefitinib-resistant NSCLC. The expression of circ_0014235, microRNA-146b-5p (miR-146b-5p) and Yes1 associated transcriptional regulator (YAP) mRNA was detected by quantitative real-time PCR (qPCR). Cell viability was detected by CCK-8 assay. Cell proliferation was assessed by colony formation assay and EdU assay. Cell cycle and cell apoptosis were determined by flow cytometry assay. The expression of marker proteins, YAP protein and programmed death ligand 1 (PD-L1) protein was detected by Western blot. The putative relationship between miR-146b-5p and circ_0014235 or YAP was ensured by dual-luciferase reporter assay and RIP assay. Animal models were established to explore the role of circ_0014235 in vivo. Circ_0014235 was highly expressed in Gefitinib-resistant NSCLC cells. Circ_0014235 downregulation reduced Gefitinib IC50, inhibited cell proliferation and induced cell apoptosis and cell cycle arrest in Gefitinib-resistant NSCLC cells, while these effects were reversed by the inhibition of miR-146b-5p, a target of circ_0014235. In addition, YAP was a target gene of miR-146b-5p, and circ_0014235 relieved miR-146b-5p-mediated inhibition on YAP by targeting miR-146b-5p. MiR-146b-5p restoration-blocked Gefitinib IC50 and cell malignant behaviors were recovered by YAP overexpression. YAP positively regulated PD-L1 expression, and YAP overexpression contributes to Gefitinib IC50 and cell malignant behaviors by upregulating PD-L1. Circ_0014235 confers Gefitinib resistance and malignant behaviors in Gefitinib-resistant NSCLC by governing the miR-146b-5p/YAP/PD-L1 pathway.

Keywords: Circ_0014235; Gefitinib; NSCLC; PD-L1; YAP; miR-146b-5p.

MeSH terms

  • Animals
  • B7-H1 Antigen
  • Carcinoma, Non-Small-Cell Lung* / drug therapy
  • Carcinoma, Non-Small-Cell Lung* / genetics
  • Carcinoma, Non-Small-Cell Lung* / pathology
  • Cell Proliferation / genetics
  • ErbB Receptors
  • Gefitinib / pharmacology
  • Lung Neoplasms* / drug therapy
  • Lung Neoplasms* / genetics
  • Lung Neoplasms* / pathology
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • RNA, Circular / genetics*

Substances

  • B7-H1 Antigen
  • MicroRNAs
  • RNA, Circular
  • ErbB Receptors
  • Gefitinib

Grant support

The author(s) reported there is no funding associated with the work featured in this article.