title: RNA interference-mediated NOTCH3 knockdown induces phenotype switching of vascular smooth muscle cells in vitro.
abstract: Notch3 plays an important role in differentiation, migration and signal transduction of vascular smooth muscle cells (VSMCs). In this study, we used RNA interference (RNAi) technique to investigate the effect of knocking down the expression of the NOTCH3 gene in VSMCs on the phenotype determination under pathologic status. Real-time PCR and Western Blot experiments verified the expression levels of Notch3 mRNA and protein were reduced more than 40% and 50% in the NOTCH3 siRNA group. When the expression of Notch3 was decreased, the proliferation, apoptosis and immigration of VSMCs were enhanced compared to control groups (P < 0.01). NOTCH3 siRNA VSMCs observed using confocal microscopy showed abnormal nuclear configuration, a disorganized actin filament system, polygonal cell shapes, and decreasing cell sizes. Additionally, knocking down the expression of NOTCH3 may evoke the CASR and FAK expression. In Conclusion, interfering with the expression of NOTCH3 causes VSMCs to exhibit an intermediate phenotype. CaSR and FAK may be involved in the Notch3 signaling pathway.
authors.affiliation: Department of Neurology, Military General Hospital of Beijing PLA Beijing, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Liu
authors.affiliation: Department of Neurology, Military General Hospital of Beijing PLA Beijing, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Neurology, Military General Hospital of Beijing PLA Beijing, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Neurology, Military General Hospital of Beijing PLA Beijing, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: Reproductive Medicine and Genetics, Military General Hospital of Beijing PLA Beijing 100700, China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: An
authors.affiliation: Department of Neurology, Military General Hospital of Beijing PLA Beijing, China.
authors.forename: Guangming
authors.initials: G
authors.lastname: Zhu
keywords: Notch3; RNAi; VSMCs; phenotype switch