Thymidine phoshorylase as a target for antiangiogenesis treatment.
Thymidine phosphorylase (TP) has emerged as a promising target for antiangiogenesis treatment of cancer. Angiogenesis, the formation of blood vessels, is essential for tumors to grow in order to be supplied with nutrients and oxygen. The association of TP with angiogenesis was demonstrated in several clinical studies in various tissue types. It has been postulated that the angiogenic effect of TP is related to its enzymatic activity, which catalyzes the breakdown of thymidine to thymine and deoxyribose-1-phosphate (dR-1-P). The latter, in its parent form or in its sugar form, deoxyribose, may play a role in angiogenesis. It may interfere in cellular energy metabolism or be substrate in a chemical reaction generating reactive oxygen species. L-deoxyribose and a specific TP inhibitor, TPI, can reverse these effects, supporting the role of the enzymatic reaction and that of the sugar. Although TP is usually high in the tumor, we also observed a high expression in tumor-associated stromal cells and macrophages. In order to elucidate the mechanism of TP induced angiogenesis we have investigated the association of TP with angiogenesis, the effect of thymidine and its metabolites on angiogenic parameters (e.g. invasion), the modulation by TPI, the formation and retention of the sugar metabolites of thymidine, and the potential signalling pathways involved in the angiogenic process. We used cell lines without/low TP expression (Colo320 and RT112) and TP transfected variants (Colo320TP1 and RT112/TP). Intrinsic TP expression in cancer cells did not stimulate these cells to invade more. On the other hand, Colo320 and Colo320TP1 cells could attract endothelial cells to a high extent, but Colo320TP1 did not attract them to a higher extent. RT112/TP cells attracted more endothelial cells than RT112 (2 fold). The difference between the RT112's and Colo320's may be related to different formation of sugars. Exposure of tumor cells to thymidine resulted in a rapid formation of dR-1-P, which was rapidly degraded to deoxyribose and further metabolized to other sugar derivatives. Of the possible sugars that can be produced by the conversion of TdR, dR-5-P seems to accumulate the most. dR accumulated 3 fold higher extent in RT112/TP than in Colo320/TP1 cells. dR could be converted to advanced glycation endproducts (AGE), however this was to a lower extent than ribose. Thymidine also induced several signalling pathways in the cells, involved in migration and invasion, such as the Focal adhesion kinase (FAK), which subsequently stimulated p70/S6 phosphorylation. The latter is a downstream kinase of rapamycin and its phosphorylation is inhibited by rapamycin, an mTOR inhibitor. The association between rapamycin and TP was shown by the protection by thymidine of rapamycin induced cytotoxicity, while TPI inhibited the effect of thymidine addition. These studies clearly show a mechanistic link between TP, signalling pathways, and cell migration.
Peters
G J
GJ
Department of Medical Oncology, VU University Medical Center, Amsterdam, the Netherlands. gj.peters@vumc.nl
Bijnsdorp
I V
IV
Fukushima
M
M
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D006801:Humans; D009369:Neoplasms / Q000201:enzymology / Q000378:metabolism; D015398:Signal Transduction; D013939:Thymidine Phosphorylase / Q000037:antagonists & inhibitors*
D020533:Angiogenesis Inhibitors; D013939:Thymidine Phosphorylase