CENPA-driven transcriptional activation of ECT2 enhances EGFR inhibitor resistance in lung adenocarcinoma

This research aims to uncover the molecular mechanisms by which centromere protein A (CENPA) and epithelial cell transforming 2 (ECT2) contribute to EGFR Inhibitor resistance in lung adenocarcinoma (LUAD). Differentially expressed genes (DEGs) related to EGFR Inhibitor resistance in LUAD were identified through analysis of the GSE153183 dataset. RT-qPCR and Western blot (WB) were performed to detect CENPA expression in osimertinib-resistant cells. A osimertinib-resistant cell line was developed by exposure to increasing concentrations of commercially obtained Osimertinib, and the drug sensitivity was evaluated using the CCK-8 assay. CENPA knockdown was achieved via lentiviral delivery, and its effects on cell proliferation, Apoptosis, migration, and invasion were evaluated. The transcriptional regulation of ECT2 by CENPA was confirmed through dual-luciferase reporter assays and chromatin immunoprecipitation (ChIP). ECT2 was exogenously overexpressed in CENPA-silenced cells, and its effects on EGFR Inhibitor resistance, epithelial-mesenchymal transition (EMT), and Cancer stemness transformation were assessed. In vivo experiments were conducted to validate the findings. A marked upregulation of CENPA expression in osimertinib-resistant LUAD cells was detected. Silencing CENPA markedly enhanced EGFR-TKI Sensitivity, suppressed colony formation, increased Apoptosis, and diminished migration and invasion. Mechanistically, CENPA promoted the expression of ECT2 via transcriptional regulation. Both in vitro and in vivo assays demonstrated that ECT2 overexpression counteracted the chemosensitizing effects of CENPA knockdown and mitigated epithelial-mesenchymal transition (EMT) and Cancer stemness transformation in the osimertinib-resistant cells. CENPA promotes EGFR Inhibitor resistance in LUAD by upregulating ECT2, which facilitates EMT and stemness transformation. Disruption of the CENPA-ECT2 pathway emerges as a new mechanism underlying EGFR Inhibitor resistance, providing promising targets for therapeutic intervention in LUAD.

CENPA; ECT2; EGFR inhibitor resistance; Lung cancer.