Author + information
The abnormal proliferation and migration of vascular smooth muscle cell (VSMC) is an important factor of vascular pathologies, such as atherosclerosis and restenosis. The study of Daxx on the regulation of VSMCs was in favor of revealing new drug targets, providing a theoretical basis for the prevention of atherosclerosis.
Firstly, we were used the lipsomal transfections take the plasmid of pCDNA3.1(+), pCDNA3.1- Daxx(+) into VSMCs to established the stable cell lines.the expression of Daxx were detected by RT- PCR and western blotting respectively. MTT and flow cytometry were respectively used to determine cellular vitality and cell cycle. Cell migration was detected by scratch wound assay. RT-PCR and western bloting were used to detect the expression of PTEN mRNA and some protein, respectively.
VSMCs treated with PDGF 20ng/ml for 24h, the cell viability, the sphase of cell cycle and the migration of VSMCs were significantly increased and the expression of Daxx protein was decreased. After transfected the Daxx gene in VSMCs, the cell viability, the sphase of cell cycle and the migration of VSMCs induced by PDGF were significantly decreased, But the shRNA-Daxx was plays an opposite role in it.Furthermore,Daxx could up-regulation the expression of PTEN gene and protein, down-regulation the expression of NF-kBp65 and p-Akt protein. When we silented PTEN gene in the Daxx gene-transfected VSMC, the cell proliferation and migration were significantly increased, which also increased the activation of NF-kB and Akt
Daxx inhibits the VSMC proliferation might though the PTEN/Akt/NF-kB signaling pathway.