Background Angiogenesis, a multi-steps process, involves interactions between cancer cells and their surrounding microenvironment. Interaction between cancer cells with these surrounding cells had been reported to be able to induce expression of angiogenesis factors or inhibitors. Aims To investigate the paracrine regulation of the important angiogenic factors (such as COX-1, COX-2, IL-8, VEGF, VEGF-C, VEGF-D, MMP-9, ICAM, NCAM, VCAM, E-Cadherin) expression, the gene expression profiles and biological phenotype change in vascular endothelial cells after interaction with lung cancer cells. Materials and Methods We use the indirect and transwell co-culture model to investigate the paracrine regulation of the important angiogenic factors (such as COX-1, COX-2, IL-8, VEGF, VEGF-C, VEGF-D, MMP-9, ICAM, NCAM, VCAM, E-Cadherin) mRNAs expression (by real-time quantitative RT-PCR) in human umbilical vein endothelial cells (HUVEC) after interaction with human lung cancer cells (CL1-5, adenocarcinoma). We also used an oligo- microarray system containing 31,000 human gene (Affymatrix array, new human gene 2.0) to perform a genome-wide analysis of the change of the gene expression profile in HUVEC after co-culture with CL1-5 lung cancer cells. In addition, we evaluate the cell proliferation, migration, invasion and other biologic characteristics changes of HUVEC after interaction with CL1-5 lung cancer cell. Results The results showed that after interaction with CL1-5, the expression of IL-8 mRNA increased about 22.6 fold, COX-2 mRNA increased 3.2 fold, ICAM mRNA increased 10.1 fold and VCAM mRNA increased 8.2 fold in the HUVEC cells (p<0.05); and other angiogenic factors showed no significant changes. By using the oligo- microarray we identify several genes that are regulated (more than 4 fold) in endothelial cells when human umbilical vein endothelial cells (HUVECs) interacts with CL1-5: there are 35 genes significantly up-regulated and 6 genes significantly down-regulated in endothelial cells. These genes expression changes involves signal transduction pathways of focal adhesion, regulation of actin cytoskeleton, Wnt signaling pathway, Jak-STAT signaling pathway, apoptosis, TGF-beta signaling pathway, tight junction, MAPK signaling pathway, and phosphatidylinositol signaling system. The biological characteristics changes as follows: no significant change in cell proliferation, significant decrease in cell migration and no significant change of cell invasion capacity of HUVEC after interaction with CL1-5 cells. Other biological properties (such as apoptosis, focal adhesion and cytoskeletal rearrangement) were also evaluated. Conclusion We concluded that HUVEC and cancer cells interaction may regulate gene expression in endothelial cells, and change the biologic characteristic of HUVEC after interaction with lung cancer cells.
[Proc Amer Assoc Cancer Res, Volume 47, 2006]