Phenotype Switch of Vascular Smooth Muscle Cells After siRNA Silencing of Filamin

Abstract

Filamin (FLN) plays an important role in differentiation, migration, and signal transduction in vascular smooth muscle cells (VSMCs). We hypothesized that suppression of FLN expression would inhibit differentiation and migration of VSMCs, and interrupt the phenotype switch of these cells. We designed and tested different shRNA sequences to silence FLN expression. The degree of silencing was assessed at mRNA (qPCR) and protein (Western blot) levels. Two sequences, FL1 and FL2, lead to the most efficient FLN silencing. Subsequent experiments were conducted using the FL1 shRNA. Cells with silenced FLN were exposed to ox-LDL, and cell phenotypic changes (cell proliferation, cell cycle, apoptosis, and morphologic changes of cytoskeleton) were evaluated. When FLN was silenced, the phenotype switch of VSMCs exposed to ox-LDL was attenuated. Further, the injury to the cytoskeleton was less prominent in the FLN-silenced cells. To conclude, RNA silencing of FLN decreases the phenotype switch of VSMCs into a pathologic state. FLN silencing could be useful in treating atherosclerosis at genetic level.