Effect of the Short Hairpin (shRNA)-SR-PSOX on Smooth Muscle Cells Migration

To investigate the effect of shRNA-SR-PSOX (Scavenger receptor binds phosphatidylserine and oxidized lipoprotein) on smooth muscle cells (SMC) migration, we designed and synthesised the shRNA oligonucleotide targeted human SR-PSOX gene, and constructed the shRNA lentiviral expression system in vitro to infect HepG2 cell line, and the silencing efficiency was assayed by real-time reverse transcription-polymerase chain reaction, immunofluorescence staining, Western blotting and Transwell chambers. The reverse transcription-polymerase chain reaction (RT-PCR) results showed that the potent inhibition of SR-PSOX expression could reach 80%, and it was specific to the SR-PSOX -derived shRNA, not the Caveolin-1-derived shRNA (negative shRNA control). Indirect immunofluorescence and Western blot results showed the SR-PSOX expression decreased obviously in SR-PSOX-shRNA transfected group compared with control group. In addition, transwell chambers results indicated that these SR-PSOX over-expressing cells showed ~3 fold increased migration of SMCs compared with normal SMC cells or siRNA cells. Red oil staining found that SR-PSOX over expressed SMCs had more lipid droplets, while lipid droplets in siRNA-SR-PSOX vector groups decreased significantly. All these data indicated that blockage and downregulation SR-PSOX expression is a potential therapeutic target to prevent inflammatory AS damage.