NEDD9 inhibits ox-LDL-induced endothelial anoikis to attenuate endothelial cell injury

Objective This study aims to explore the action of the anoikis gene in the vascular endothelial cell injury.Methods The set of genes associated with anoikis was utilized for conducting enrichment analysis on the Gene Expression Omnibus (GEO: GSE100927) dataset to identify the common genes linked to Atherosclerosis. Subsequently, the expression levels and pathway enrichment of anoikis genes in the GSE100927 dataset were examined. The LASSO method was employed for reducing dimensionality in modeling to identify genes correlated with Atherosclerosis and to establish an anoikis score. The validation of the expression of neural precursor cell-expressed developmentally down-regulated 9 (NEDD9), FOSB, and ERCC1 in ox-LDL-induced bEnd.3 cells was carried out using RT-qPCR. Atherosclerotic mice were established to evaluate NEDD9 expression. The impact of overexpressing or silencing NEDD9 on anoikis in ox-LDL or detachment-induced bEnd.3 cells or Human Umbilical Vein Endothelial Cells (HUVEC) was assessed through CCK-8, EdU, and flow cytometry assays.Results Based on anoikis gene analysis, NFIL3, NR4A3, ADAMTS4, NEDD9, STX17-AS1, and CSF3 were found to be under-expressed, while FOSB and ERCC1 were found to be over-expressed in the atherosclerosis group compared to the normal group. LASSO regression analysis yielded an anoikis score = -9.522e-01 × NFIL3-3.410 × NEDD9 + 2.728e-01 × ADAMTS4 + 1.178 × FOSB +5.896e-15 × ERCC1 + 1.558e+01. Compared with the blank group, NEDD9, FOSB, and ERCC1 were under-expressed in the ox-LDL intervention group. NEDD9 protein was significantly decreased in the arterial tissues of atherosclerotic mice. si-NEDD9 promoted an increase in ROS and apoptosis levels in the bEnd.3 cells intervened by ox-LDL. Transfection with oe-NEDD9 increased the viability of bEnd.3 cells, upregulated the ITGB1 and E-cadherin proteins level, while decreasing ROS and apoptosis levels induced by the ox-LDL and detachment. Overexpression of NEDD9 also reduced HUVEC anoikis induced by detachment.Conclusion A preliminary atherosclerotic anoikis model was established to predict endothelial cell injury. Overexpression of NEDD9 attenuated ox-LDL and detachment-induced endothelial cell anoikis in vitro, suggesting a potential protective role.