Dendritic cell-derived lncRNA in patients with acute coronary syndrome

Abstract

BackgroundLong non-coding RNA (lncRNA) and dendritic cells (DC) play an important role in the occurrence and development of acute coronary syndrome (ACS). However, the role and mechanism of DC-derived lncRNA in patients with ACS still remains understudied.ObjectiveTo investigate the expression difference and role of lncRNA in monocyte-derived DC(moDC) in patients with acute coronary syndrome(ACS).Methods and ResultsThe subjects were divided into 4 groups: (1) healthy control group (CON), (2) unstable angina group (UA), (3) non-ST elevation myocardial infarction group (NSEMI or NST) and (4) ST elevation myocardial infarction group (STEMI or ST). Peripheral blood was extracted and isolated to obtain mononuclear cells (PBMC). After induction into moDC, morphology was observed by electron microscope and identified by flow cytometry (FCM) (CD80, CD86, CD11c, CD14, HLA-DR), and morphological and functional differences were compared. The differentially expressed lncRNA were screened by gene sequencing, and the biological functions of which were predicted by GO and KEGG and the correlation between candidate lncRNA and target genes was analyzed. The expression levels of markers, signaling pathways, inflammatory factors and target gene were performed by FCM, RT-PCR, WB and ELISA after the candidate lncRNA were over-expressed or silenced. Electron microscope showed that the cells were suspended cells of dendritic pseudopodia. FCM showed that the expression levels of DC markers CD11c (99.02±0.12%) and HLA-DR (99.32±0.08%) were high, while the expression levels of CD80 (24.27±0.99%) and CD86 (32.38±0.59%) were low. The number of differentially expressed lncRNA screened by gene sequencing was as follows: CON vs UA, 3; CON vs. NST, 49; CON vs ST, 35; UA vs. NST, 115; UA vs ST, 113; ST vs NST, 4. The results of FCM, RT-PCR, WB and ELISA showed that the expression levels of markers (CD80, CD86, HLA-DR), signaling pathways (p-PI3K, p-AKT), inflammatory factors (IL-6 and IL-12p70) and target gene (CCL14) were increased after over-expression of CCL15-CCL14; On the contrary, its expression level was decreased.ConclusionsThere are differences in moDC morphology and function and lncRNA expression in different types of ACS patients, and the differentially expressed lncRNA (CCL15-CCL14) regulates the function of moDC.