Research Article
Knockdown of AAMP Impacts on HECV Cell Functions In Vitro and Affects the Expression of VE-Cadherin
Yukun Yin1,2,3, Andrew J Sanders1, Li Feng2 and Wen G Jiang1*1Cardiff University-Peking University Cancer Institute, Cardiff University School of Medicine, Heath Park, Cardiff, UK
2Department of Oncology, Wangjing Hospital of China Academy of Traditional Chinese Medicine, Hua Jia Di Street, ChaoYang District, Beijing 100102, P.R. China
3China Academy of Traditional Chinese Medicine, Nan Xio Jie, Dong Cheng District, Beijing, 100700, P.R. China
- *Corresponding Author:
- Wen G Jiang
Cardiff University-Peking University Cancer Institute
Cardiff University School of Medicine, Cardiff, CF14 4XN, UK
Tel: 44 29 2068 7065
Fax: 44 29 2076 1623
E-mail: [email protected]
Received Date: January 17, 2014; Accepted Date: February 21, 2014; Published Date: February 28, 2014
Citation:Yin Y, Sanders AJ, Feng L, Jiang WG (2014) Knockdown of AAMP Impacts on HECV Cell Functions In Vitro and Affects the Expression of VECadherin. Angiol 2:125. doi: 10.4172/2329-9495.1000125
Copyright: © 2014 Yin Y, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Background: Angio-Associated Migratory Cell Protein (AAMP) is a 52 kDa protein expressed in a variety of human cell lines. Previous studies have demonstrated that AAMP is involved in endothelial cell adhesion, migration, and tube formation and plays roles in signalling pathways, such as RhoA-ROCK
Methods: In this study, we knocked down the expression of AAMP in HECV cells using hammerhead ribozymes and assessed the influence of AAMP on endothelial cells. In order to explore potential mechanisms, Wnt/β catenin inhibitors (FH535 or IWP2) were also used in a number of the functional assays and the immunofluorescent staining patterns of several key molecules were explored.
Results: Knockdown of AAMP expression was observed in the HECV cell line following transfection with the ribozyme transgene. Cellular migration and attachment, assessed using ECIS methods, and tubule formation were significantly inhibited by the knockdown of AAMP. Additionally, cell growth and cell matrix adhesion was also substantially reduced following AAMP knockdown, though this did not reach significance. Cell aggregation levels showed no statistical difference between AAMP knockdown cells and control cells. Interestingly, immunofluorescence staining showed AAMP knockdown cells had a reduced expression of VE-cadherin. Some overlap of function was seen between AAMP knockdown and the FH535 inhibitor.
Conclusion: AAMP appears to influence endothelial cell migration and tubule formation and potentially, to a lesser effect, cell matrix adhesion and growth and also affects the expression of VE-cadherin. This data suggests that AAMP may play a role in angiogenesis. Assays performed with the FH535 inhibitor suggest a potential relationship between AAMP and Wnt/β catenin signalling.
