YY1-Regulated AEBP1 Drives the Progression and Inhibits Ferroptosis of Lung Adenocarcinoma

Abstract

Background: Lung adenocarcinoma (LUAD) is a severe type of lung cancer, characterized by high prevalence and mortality rate. Adipocyte enhancer binding protein 1 (AEBP1) is a critical cancer-promoting factor in several types of cancer. Nevertheless, the hidden mechanism underlying the effect of AEBP1 on LUAD has not been investigated. Methods: The mRNA and protein expressions of AEBP1 and Yin Yang 1 (YY1) were detected by reverse transcription-quantitative PCR (RT-qPCR) and western blot. By means of Cell Counting Kit-8 (CCK-8), 5-ethynyl-2′-deoxyuridine (EdU) staining, colony formation assay, transwell and wound healing assays, A549 cell viability, cell proliferation, invasion and migration were appraised. The angiogenesis of human umbilical vein endothelial cells (HUVECs) was evaluated with tube formation assay. Western blot was used to detect proteins associated with epithelial-mesenchymal transition (EMT) and ferroptosis. Thiobarbituric acid-reactive substances (TBARS) production rate and Fe²⁺ levels were measured using the corresponding kits. Reactive oxygen species (ROS) activity was detected by 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C11-BODIPY(581/591)). Luciferase reporter assay was used to detect AEBP1 promoter activity and chromatin immunoprecipitation was performed to estimate the binding ability of YY1 and AEBP1 promoter. Results: AEBP1 silencing inhibited the proliferation, metastasis and angiogenesis whereas induced ferroptosis in LUAD cells (p < 0.001), which were all reversed by YY1 (p < 0.05 or p < 0.01 or p < 0.001). Besides, YY1 was found to promote AEBP1 transcription (p < 0.001). Conclusion: Collectively, YY1-regulated AEBP1 could facilitate LUAD advancement and inhibit ferroptosis.