Overexpression of Neural Precursor Cell Expressed Developmentally Downregulated 9 (NEDD9) reduces ox-LDL-induced Anoikis in atherosclerotic vascular endothelial cells.

OBJECTIVE: This study purposes to explore the action of the Anoikis gene in vascular endothelial cell injury, explore diagnostic biomarkers, and provide new insights into potential molecular mechanisms, as well as offer a new perspective for disease detection and treatment. METHODS: The Anoikis gene set was used for enrichment analysis on the Gene Expression Omnibus (GEO: GSE100927) dataset, to identify the intersection genes related to Atherosclerosis. Further, the expression and pathway enrichment of Anoikis genes in GSE100927 was investigated. The Least Absolute Shrinkage and Selection Operator (LASSO) method for dimensionality reduction modeling was employed to obtain Atherosclerosis-related genes and construct Anoikis score. The NEDD9, FOSB, and ERCC1 expression in ox-LDL-induced the Bend.3 cells was validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Overexpression or silencing NEDD9 on Anoikis in ox-LDL and detachment-induced the Bend.3 cells was analyzed by using Cell Counting Kit-8 (CCK8), 5-Ethynyl-2'-deoxyuridine (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 = -9.522(e-01) × NFIL3 - 3.410 × NEDD9 + 2.728(e-01) × ADAMTS4 + 1.178 × FOSB + 5.896(e-15) × ERCC1 + 1.558(e+01). Compared with the blank group, NEDD9, FOSB, and ERCC1 were under-expressed in the ox-LDL intervention group. si-NEDD9 promoted an increase in reactive oxygen species (ROS) and apoptosis levels in the Bend.3 cells intervened by ox-LDL. Transfection with oe-NEDD9 increased the viability of Bend.3 cells induced by the ox-LDL and detachment, while decreasing ROS and apoptosis levels. CONCLUSION: This study developed a reliable atherosclerotic Anoikis model for predicting endothelial cell injury. During Anoikis genes, the overexpression of NEDD9 reduces ox-LDL and detachment-induced endothelial cell Anoikis.