Background: Platelet-derived endothelial cell growth factor (PD-ECGF), whose expression is increased in several cancers, is an endothelial cell mitogen and has chemotactic activity in vitro and angiogenic activity in vivo. Tumors with high PD-ECGF expression tend to have frequent lymph node metastasis and are associated with poor outcome.
Methods: We screened genes transduced by PD-ECGF transfection to the colon cancer cell line DLD-1 by using a complementary DNA microarray. Cell motility was evaluated by in vitro migration assay. Actin fiber polymerization was visualized by immunofluorescent detection of phalloidin.
Results: Rho-associated coiled-coil domain kinase (ROCK1) was found to be significantly overexpressed in PD-ECGF transfectants compared with mock cells. PD-ECGF transfectants showed higher cell motility than mock cells. The parental cell, DLD-1, with recombinant PD-ECGF showed higher cell motility than that without recombinant PD-ECGF, in which motility was blocked by the neutralizing antibody of PD-ECGF or Y-27632, a specific inhibitor of ROCK1. Moreover, the actin fiber polymerization, which is a marker of activation of ROCK1, was higher in PD-ECGF transfectants than in mock cells.
Conclusions: PD-ECGF expression may be associated with cancer cell migration via activation of ROCK1. This may explain one mechanism by which tumors with high expression of PD-ECGF show aggressive behavior.
PD-ECGF ROCK1 Cell motility cDNA microarray
This is a preview of subscription content, log in to check access
Mimori K, Ueo H, Shirasaka C, et al. Up-regulated pyrimidine nucleoside phosphorylase in breast carcinoma correlates with lymph node metastasis. Ann Oncol 1999; 10: 111–3.PubMedGoogle Scholar
Yamagata M, Mori M, Mimori K, et al. Expression of pyrimidine nucleoside phosphorylase mRNA plays an important role in the prognosis of patients with oesophageal cancer. Br J Cancer 1999; 79: 565–9.PubMedGoogle Scholar
Matsuura T, Kuratate I, Teramachi K, Osaki M, Fukuda Y, Ito H. Thymidine phosphorylase expression is associated with both increase of intratumoral microvessels and decrease of apoptosis in human colorectal carcinomas. Cancer Res 1999; 59: 5037–40.PubMedGoogle Scholar
Takao S, Akiyama S, Nakajo A, et al. Suppression of metastasis by thymidine phosphorylase inhibitor. Cancer Res 2000; 60: 5345–8.PubMedGoogle Scholar
Miyadera K, Sumizawa T, Haraguchi M, et al. Role of thymidine phosphorylase activity in the angiogenic effect of platelet derived endothelial cell growth factor/thymidine phosphorylase. Cancer Res 1995; 55: 1687–90.PubMedGoogle Scholar
Nishida M, Hino A, Mori K, Matsumoto T, Yoshikubo T, Ishitsuka H. Preparation of anti-human thymidine phosphorylase monoclonal antibodies useful for detecting the enzyme levels in tumor tissues. Biol Pharm Bull 1996; 19: 1407–11.PubMedGoogle Scholar
Mori M, Mimori K, Sadanaga N, et al. Prognostic impact of tissue inhibitor of matrix metalloproteinase-1 in esophageal carcinoma. Int J Cancer 2000; 88: 575–8.PubMedGoogle Scholar
Rubin E, Gill DM, Boquet P, Popoff MR. Functional modification of a 21-kilodalton G protein when ADP-ribosylated by exoenzyme C3 of Clostridium botulinum. Mol Cell Biol 1988; 8: 418–26.PubMedGoogle Scholar
Just I, Selzer J, Eichel-Streiber C, Aktories K. The low molecular mass GTP-binding protein Rho is affected by toxin A from Clostridium difficile. J Clin Invest 1995; 95: 1026–31.CrossRefPubMedGoogle Scholar
Itoh K, Yoshioka K, Akedo H, Uehata M, Ishizaki T, Narumiya S. An essential part for Rho-associated kinase in the transcellular invasion of tumor cells. Nat Med 1999; 5: 221–5.PubMedGoogle Scholar
Moghaddam A, Zhang HT, Fan TP, et al. Thymidine phosphorylase is angiogenic and promotes tumor growth. Proc Natl Acad Sci U S A 1995; 92: 998–1002.PubMedGoogle Scholar
Klein RS, Lenzi M, Lim TH, Hotchkiss KA, Wilson P, Schwartz EL. Novel 6-substituted uracil analogs as inhibitors of the angiogenic actions of thymidine phosphorylase. Biochem Pharmacol 2001; 62: 1257–63.PubMedGoogle Scholar
Matsumoto Y, Tanaka K, Harimaya K, Nakatani F, Matsuda S, Iwamoto Y. Small GTP-binding protein, Rho, both increased and decreased cellular motility, activation of matrix metalloproteinase 2 and invasion of human osteosarcoma cells. Jpn J Cancer Res 2001; 92: 429–38.PubMedGoogle Scholar
Bhowmick NA, Ghiassi M, Bakin A, et al. Transforming growth factor-beta1 mediates epithelial to mesenchymal transdifferentiation through a RhoA-dependent mechanism. Mol Biol Cell 2001; 12: 27–36.PubMedGoogle Scholar
Sahai E, Olson FM, Marshall JC. Cross-talk between Ras and Rho signaling pathways in transformation favours proliferation and increased motility. EMBO J 2001; 20: 755–66.PubMedGoogle Scholar
Ayaki M, Mukai M, Imamura F, et al. Cooperation of fibronectin with lysophosphatidic acid induces motility and transcellular migration of rat ascites hepatoma cells. Biochim Biophys Acta 2000; 1495: 40–50.PubMedGoogle Scholar
Kanyama H, Tomita N, Yamano T, et al. Enhancement of the anti-tumor effect of 5′-deoxy-5-fluorouridine by transfection of thymidine phosphorylase gene into human colon cancer cells. Jpn J Cancer Res 1999; 90: 454–9.PubMedGoogle Scholar
Koizumi W, Saigenji K, Nakamaru N, Okayasu I, Kurihara M. Prediction of response to 5′-deoxy-5-fluorouridine (5′-DFUR) in patients with inoperable advanced gastric cancer by immunostaining of thymidine phosphorylase/platelet-derived endothelial cell growth factor. Oncology 1999; 56: 215–22.PubMedGoogle Scholar
Ishikawa T, Sekiguchi F, Fukase Y, Sawada N, Ishitsuka H. Positive correlation between the efficacy of capecitabine and doxifluridine and the ratio of thymidine phosphorylase to dihydropyrimidine dehydrogenase activities in tumors in human cancer xenografts. Cancer Res 1998; 58: 685–90.PubMedGoogle Scholar