| Angiogenesis is recognized as a fundamental process and an essential componentof tumour growth and metastasis. Anti-angiogenesis has become a promising strategyfor the treatment of cancer and anti-angiogenic agents can be developed as novelanticancer agents. The aim of this study was to investigate the inhibitory effect ofrecombinant CS5931polypeptide on angiogenesis both in vitro and in vivo using theHUVEC and the zebrafish model, respectively and the possible anti-angiogenesismechanism of recombinant CS5931polypeptide was also presented.CS5931is a novel polypeptide isolated from Ciona Savignyi with potent anticanceractivities and its molecular weight is5931Da. Recombinant CS5931was expressedusing a prokaryotic expression system and also showed potent anticancer activity. Inthe present study, we first determine the effect of the recombinant polypeptide on theproliferation, migration and the capillary-like tube formation of HUVECs (HumanUmbilical Vein Endothelial Cell). The results showed that the polypeptide couldinhibite the proliferation of HUVECs, prevent the endothelial cell migration, anddestruct the tube formation of HUVECs in a concentration-dependent manner.To further study the mechanism underlying its anti-angiogenic activity, theexpression of VEGF (Vascular Endothelial Growth Factor) and MMP-2/-9(MatrixMetalloproleinase-2/-9) were determined using Western blotting and gelatinzymography. These results revealed that recombinant CS5931polypeptide couldeliminate VEGF production without any effect on the mRNA level, and thepolypeptide could reduce the secretion and expression of MMP-2/9, as well as their mRNA levels.We then checked the anti-angiogenic activity of the polypeptide in vivo usingzebrafish model. After treatment with certain concentrations of recombinant CS5931polypeptide, the angiogenic defects of zebrafish embryos were observed using adissecting stereomicroscope. The results showed that incubation with30μg/mLCS5931for48h clearly caused pericardial edema, heart weakness and reduced bloodflow rates in the zebrafish embroys; alkaline phosphatase staining indicated thatrecombinant CS5931polypeptide prevented SIV formation in zebrafish embryossignificantly in a dose-dependent relationship.In summary, we found that recombinant CS5931polypeptide has anti-angiogenicactivity both in vitro and in vivo models and we revealed that recombinant CS5931polypeptide could inhibit VEGF and MMPs, which maybe contribute to theanti-angiogensis. |
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