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Identification of tumour-induced changes in endothelial cell surface protein expression: an in vitro model
1University of Nottingham Laboratory of Molecular Oncology, CRC Academic Department of Clinical Oncology, City Hospital, Hucknall Rd, NG5 1PB, Nottingham, UK. peter.hewett@nott.ac.uk
Abstract:
The selective destruction of the supporting vasculature of tumours has been proposed as a means of therapy. Fundamental to this approach is the identification of suitable targets on tumour-endothelium. To detect proteins that may be up-regulated on the luminal (apical) surface of tumour-associated endothelium confluent endothelial cells were examined following incubation with tumour cell conditioned medium (TCM) from, or co-culture with, a range of breast carcinoma and small cell lung carcinoma (SCLC) cell lines. Exposed endothelial membrane proteins were labelled with sulpho-NHS-biotin and detected by enhanced chemiluminescence following two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and western blotting. TCM induced varying levels of proliferative activity in endothelial cells; generally breast TCM contained greater mitogenic activity than SCLC TCM. Exposure of human breast and lung microvascular, and umbilical vein endothelial cells to soluble tumour cell factors from several breast cancer and SCLC cells lines produced similar changes in luminal protein profiles: Breast cancer cells and in particular the MDA-MB-231 cell line induced the most pronounced changes. The expression of six proteins was altered consistently on endothelial cells stimulated with soluble tumour cell factors. However, similar changes were observed following incubation with ECGS suggesting that they were related to endothelial cell proliferation per se. As these proteins were altered in breast and lung microvascular, and umbilical vein endothelial cells stimulated by a variety of breast cancer and SCLC cell lines they support the potentially broad applicability of anti-vascular approaches targeted at the endothelium.
Insights
Researchers identified proteins on tumor-associated endothelial cells for anti-vascular cancer therapy. Tumor cell factors alter endothelial cell protein expression, suggesting broad applicability for targeting tumor vasculature.
Area of Science:
- Oncology
- Vascular Biology
- Proteomics
Background:
- Targeting tumor vasculature is a promising therapeutic strategy.
- Identifying specific targets on tumor-associated endothelium is crucial for this approach.
- Tumor cell-derived factors can influence endothelial cell behavior and protein expression.
Purpose of the Study:
- To identify proteins upregulated on the luminal surface of tumor-associated endothelial cells.
- To investigate the effects of tumor cell-conditioned medium (TCM) and co-culture on endothelial cell protein profiles.
- To assess the potential of these identified proteins as targets for anti-vascular therapies.
Main Methods:
- Endothelial cells were incubated with TCM or co-cultured with breast carcinoma and small cell lung carcinoma (SCLC) cell lines.
- Lumincal endothelial membrane proteins were labeled with sulpho-NHS-biotin.
- Proteins were detected using enhanced chemiluminescence after two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and western blotting.
Main Results:
- Tumor cell-conditioned medium induced varying levels of endothelial cell proliferation, with breast cancer TCM showing higher mitogenic activity.
- Exposure to soluble tumor cell factors altered luminal protein profiles in human breast and lung microvascular, and umbilical vein endothelial cells.
- Six proteins showed consistent expression changes upon stimulation with soluble tumor cell factors, though some changes were also linked to general endothelial cell proliferation.
Conclusions:
- Soluble factors from various breast cancer and SCLC cell lines consistently alter endothelial cell protein expression.
- These altered proteins are present across different endothelial cell types, indicating broad applicability.
- The findings support the potential of anti-vascular approaches targeting the tumor endothelium for cancer therapy.