NANOG adenovirus

Here, the researchers explored the expression of nanoprotein homeobox (NANOG) in thoracic aortic dissection (TAD) and its role in regulating the phenotypic switch of human aortic vascular smooth muscle cells (VSMCs). The experimental results showed that NANOG and OPN were highly expressed in the aortic wall and VSMCs in TAD, and both were accompanied by the phenotypic switch of VSMCs. Overexpression of NANOG induced upregulation of matrix metalloproteinase 2, a synthetic marker of VSMCs, and downregulation of contractile markers α-smooth muscle actin and smooth muscle 22α. Overexpression of NANOG also enhanced the proliferation, migration and anti-apoptosis ability of VSMCs. The results also showed that these functions of NANOG were achieved through OPN, and NANOG directly upregulated OPN by binding to its promoter region.

Smooth muscle cells (VSMCs) were isolated from control samples and transfected with NANOG overexpression adenovirus (Ad-NANOG) to investigate the effects of NANOG on osteopontin (OPN) expression and smooth muscle cell phenotype. Transfection effects were detected by fluorescence observation, qRT-PCR, and Western blot (Figure 1A, B, and D). The expression level of NANOG protein relative to β-actin was measured, and the results were similar to those in TAD VSMCs. The researchers first evaluated the expression changes of related genes at the mRNA level compared with VSMCs transfected with Ad-GFP. After 48 hours of transfection, the mRNA expression level of OPN in Ad-NANOG VSMCs increased to 2.30 ± 0.20 (Figure 1C). In Ad-NANOG VSMCs, the VSMC synthesis marker MMP2 increased to 11.17±2.98, but the VSMC contraction markers α-SMA and SM22α decreased to 0.71±0.13 and 0.41±0.12, respectively (Figure 1F-H). Western blot results confirmed these changes at the protein level (Figure 1E, I-K).

Figure 1. Overexpression of NANOG up-regulated the expression of OPN and VSMCs synthetic MMP2, whereas the expression of VSMCs contractile markers α-SMA and SM22α were down-regulated. (An Z, et al., 2017)