Analysis of cardiovascular disease-related NF-κB-regulated genes and microRNAs in TNFα-treated primary mouse vascular endothelial cells

Activated nuclear factor-κB(NF-κB)plays an important role in the development of cardiovascular disease(CVD)through its regulated genes and microRNAs(miRNAs).However,the gene regulation profile remains unclear.In this study,primary mouse vascular endothelial cells(pMVECs)were employed to detect CVD-related NF-κB-regulated genes and miRNAs.Genechip assay identified 77 NF-κB-regulated genes,including 45 upregulated and 32 downregulated genes,in tumor necrosis factorα(TNFα)-treated pMVECs.Ten of these genes were also found to be regulated by NF-κB in TNFα-treated He La cells.Quantitative real-time PCR(q RT-PCR)assay confirmed the upregulation of Egr1,Tnf,and Btg2 by NF-κB in the TNFα-treated p MVECs.The functional annotation revealed that many NF-κB-regulated genes identified in pMVECs were clustered into classical NF-κB-involved biological processes.Genechip assay also identified 26 NF-κB-regulated miRNAs,of which 21 were upregulated and 5 downregulated,in the TNFα-treated pMVECs.Further analysis showed that nine of the identified genes are regulated by seven of these mi RNAs.Finally,among the identified NF-κB-regulated genes and miRNAs,5 genes and 12 miRNAs were associated with CVD by miRWalk and genetic association database analysis.Taken together,these findings show an intricate gene regulation network raised by NF-κB in TNFα-treated p MVECs.The network provides new insights for understanding the molecular mechanism underlying the progression of CVD.