Effect of p54nrb gene silencing on the endothelial cells migration of human umbilical vein and the angiogenesis in vitro / 解放军医学杂志

ABSTRACT

Objective To explore the effects of p54nrb gene silencing on the proliferation, migration of human umbilical vein endothelial cell (HUVEC) and angiogenesis in vitro. Methods HUVECs were infected by p54nrb-specific shRNA-containing lentiviruses for 24h, then the cells were cultured in media containing puromycin until stable establishment of p54nrb-silencing HUVEC cell line. The efficiency of p54nrb gene silencing technique was determined by Western blotting, and the effects of this technique on the proliferation of HUVECs was assessed by the Cell Counting Kit-8 (CCK-8) assay. The migration of p54nrbsilencing HUVECs was measured by wound healing assay and Transwell motility assay. Vessel formation assay was used to detect the angiogenesis ability of p54nrb-silencing HUVECs. Results Western blotting showed that the expression of p54nrb protein in p54nrb-silencing HUVECs decreased significantly, implying a stable establishment of p54nrb-silencing HUVEC cell line. The CCK-8 assay revealed that knockdown of p54nrb gene promoted the proliferation of HUVECs slightly. Wound healing assay and Transwell motility assay displayed that knockdown of p54nrb significantly inhibited the motility of HUVECs. Vessel formation assay showed that knockdown of p54nrb inhibited in vitro the formation of vessel-like structures of HUVEC cells. Conclusion p54nrb significantly promote the migration and angiogenesis in vitro of HUVECs, and may be an important modulin involved in angiogenesis.