Changes in microvessel endothelial cell gene expression in an in vitro human breast tumour endothelial cell model |
| |
Authors: | Peter Hewett Andrew Popplewell Helene Finney J. Clifford Murray |
| |
Affiliation: | (1) CRC Academic Department of Clinical Oncology, University of Nottingham Laboratory of Molecular Oncology, City Hospital, Hucknall Road, Nottingham, NG5 1PB, UK;(2) Celltech Chiroscience, 216 Bath Rd, Slough, SL1 4EN, UK |
| |
Abstract: | Selective targeting of tumour-endothelium has been proposed as a means of therapy. The successful exploitation of this approach will rely on the identification of suitable targets expressed specifically on the tumour-associated endothelium. In an attempt to identify novel tumour-endothelium associated targets we have used differential mRNA display to identify genes up-regulated in an in vitro breast tumour-endothelial cell culture model. Confluent monolayers of human mammary microvessel endothelial cells (HuMMEC) were incubated for 5 days with MDA-MB-231 breast adenocarcinoma cell-conditioned medium (TCM). mRNAs isolated from TCM-treated and control cells were amplified using 104 combinations of four 3′ anchored T12VN primers and 26 ‘random’ 10mers by RT-PCR and the products examined on DNA sequencing gels. Seventy-four sequences were cloned and the differential expression of five genes was confirmed using dot-blots. These were identified as procollagen type-IV, Tie-2/Tek receptor tyrosine kinase, NADH dehydrogenase subunit-6, and ferritin heavy-chain, which were up-regulated, and insulin-like growth factor binding protein-5, which was down-regulated. Increased endothelial expression of basement membrane proteins and tyrosine kinase receptors is known to occur during angiogenesis. Our data support the use of this model for further in vitro investigation of tumour angiogenesis. This revised version was published online in June 2006 with corrections to the Cover Date. |
| |
Keywords: | angiogenesis differential mRNA display endothelial cells Tie-2/Tek receptor |
本文献已被 PubMed SpringerLink 等数据库收录! |
|