We describe the effect of recombinant canstatin, the NC1 domain of the α2 chain of
Type IV collagen, on suppression of angiogenesis and lymphangiogenesis both in vitro and in vivo. Recombinant canstatin produced from stably transformed Drosophila S2 cells reduced the expression of
angiopoietin-1 in
hypoxia mimetic agent,
CoCl(2) -treated CT-26 cells. Recombinant canstatin inhibited proliferation, tube formation and migration of human
angiopoietin-1 (rhAngpt-1)-treated human umbilical vein endothelial cells (HUVEC) and lymphatic endothelial cells (LEC). Recombinant canstatin suppressed the expression of Tie-2 and vascular endothelial growth factor-3 (VEGFR-3) transcripts in rhAngpt-1-treated HUVEC and LEC, respectively. The inhibitory effect of recombinant canstatin on
tumor growth was also investigated using a heterotopic CT-26 colon
carcinoma animal (BALB/c mice) model. Recombinant canstatin reduced the final volume and weight of
tumors, and blood and lymphatic vessel densities of
tumors, which were evaluated by CD-31 and LYVE-1 immunostaining. Immunohistochemical analysis showed that recombinant canstatin dramatically reduced the expression of
angiopoietin-1 in CT-26 colon
carcinoma-induced
tumor, but not the expression of
VEGF-C. Tie-2 and
VEGFR-3 expressions were also reduced in recombinant canstatin-treated
tumors. These results indicate that recombinant canstatin has anti-tumoral activities against CT-26 colon
carcinoma cells. Recombinant canstatin reduces the expression of
angiopoietin-1 in
hypoxia-induced CT-26 cells and inhibits the angiogenic and lymphangiogenic signaling induced by
angiopoietin-1. Recombinant canstatin probably inhibits angiogenesis and lymphangiogenesis via suppression of the
integrin-dependent FAK signaling induced by
angiopoietin-1/Tie-2 and/or
VEGFR-3.