Knockdown of caveolin-1 inhibits the tubule-like formation and increases the NO release of MS1 endothelial cells

• Main Authors: Chung-Mei Chung, 鄭春梅
• Other Authors: Yat-Pang Chau
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/928sh6

碩士 === 國立陽明大學 === 解剖暨細胞生物學研究所 === 97 === The structure and activation of membrane proteins play key regulatory roles in numerous intracellular signal transduction in eukaryotic cells. Caveolae, a 50 nanometer-sized invagination of the plasma membrane, is well-known microstructure for endocytosis and transcytosis in endothelial cells. Caveolin-1 (CAV1) is a major component of caveolae. Strong evidence showed that caveolin-1 acts as a linker for eNOS and plays an important role for NO signaling in endothelial cells. In this study, we explored the functional roles of CAV1 in endothelial MS1 cells with RNA interference by using VSV-G pseudotyped lentivirus system. By shCAV1-lentiviral infection technique, we downregulated CAV1 expression in endothelial MS1 cells which express high levels of CAV1 mRNA and caveolin-1 protein. The infection of shCAV1 to MS1 cells (MS1cav1- cells) successful reduced the levels of CAV1mRNA and its protein about 46% and 45% compared with mock cells, respectively. Moreover, MS1cav1- cells have no morphological change and alteration of cell growth. However, the knock-down of CAV1 expression resulted in the decrease of angiogenic factors, (VE-cadherin and Tie-2), leading to the failure of tubule-like formation, and the downregulation of cell adhesion molecules (JAM-1、Itgb5、VCAM-1), leading to the decrease of cell adhesion. On the other hand, the knock-down of CAV1 expression in MS1cav1- cells increases the nitric oxide release and promotes the cell migration ability.