Mechanisms of aortic dissection smooth muscle cell phenotype switch

Abstract

Objective: To investigate the expression of Nanog homeobox (NANOG) in thoracic aortic dissection (TAD) and the role of NANOG in regulating human aortic vascular smooth muscle cells (VSMCs) phenotype switch.

Methods: Aortic specimens were collected from 20 patients undergoing TAD and 10 controls. VSMCs were isolated by adherent cultivation approach. The expression of NANOG, osteopontin (OPN), and VSMCs phenotype markers were determined by quantitative real-time polymerase chain reaction, Western blot, immunohistochemistry, and immunofluorescence. Cell counting, scratch wound-healing assay, Transwell migration, and apoptosis assays were used for cell function assessment. Deoxyribonucleic acid-protein binding detection was performed by chromatin immunoprecipitation.

Results: Our experiment results showed that NANOG and OPN were highly expressed in TAD aortic wall and VSMCs, both accompanying VSMCs phenotype switch. Overexpression of NANOG induced the up-regulation of VSMCs synthetic marker matrix metalloproteinase 2 and the down-regulation of VSMCs contractile markers α-smooth muscle actin and smooth muscle 22α. Overexpression of NANOG also enhanced the proliferation, migration, and antiapoptosis capabilities of VSMCs. The results also showed that these functions of NANOG was via OPN and NANOG directly up-regulated OPN by binding to its promoter region.

Conclusions: Our study suggests that NANOG is highly expressed in TAD aortic wall and VSMCs. Increased NANOG promotes VSMCs phenotype switch by directly up-regulating OPN through binding to its promoter region.

Keywords: NANOG; OPN; aortic dissection; phenotype switch; smooth muscle cell.