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Cytotoxic Effects of Transforming Growth Factor-α Conjugated with Cytotoxin Saporin on Proliferating Vascular Smooth Muscle Cells

Abstract Objective To testify the special cytotoxicity of TGF-alpha-SAP on proliferating vascular smooth muscle cells and endothelial cells. Methods Conjugation of saporin to TGF-alpha was accomplished after derivatization of saporin and TGF-alpha with N-succinimidyl-3 (2-pyridyldithio) proprionate and final purification of the conjugate was achieved within Eppendorf Centrifugal Filter Cytotoxicity assays were measured by cell count. The studies of influence of TGF-alpha-SAP on values of Thymidine and leucine incorporation into SMCs and ECs were measured by 3H-thymidine uptake and 3H-leucine uptake, respectively. and receptor competition studies of TGF-alpha-SAP are measured by adding excess TGF-alpha in SMCs exposed for TGF-SAP. Results Cytotoxicity assays testified TGF-alpha-SAP conjugate could inhibit remarkably proliferation of SMCs in culture. The values of thymidine of TGF-alpha-SAP group (10-9M and 10-7M) in comparison significantly decreased to 60.9% and 56.0% of the control group respectively, suggesting that cellular DNA synthesis obviously decreased as TGF-alpha-SAP was added. But Saporin did not affect cellular DNA synthesis at higher level. The rate of 3H-leucine incorporation of TGF-alpha-SAP group significantly decreased to 47.3% of the control group, suggesting that SMCs protein synthesis obviously decreased as TGF-alpha-SAP was added. But TGF-alpha-SAP at the same level did not affect DNA synthesis and protein synthesis of ECs compared with the control group. Conclusion The results indicated that TGF-alpha-SAP possesses the more effective cytotoxicity than Saporin and the more special citotoxicity on proliferating vascular smooth muscle cells than on proliferating endothelial cells. Key words：cytotoxicity;selective; Drug-eluting stents;proliferation;saporin