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


Article Preview

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



Edited by:

Qingzhou Xu




J. Yang et al., "Cytotoxic Effects of Transforming Growth Factor-α Conjugated with Cytotoxin Saporin on Proliferating Vascular Smooth Muscle Cells", Advanced Materials Research, Vol. 621, pp. 182-187, 2013

Online since:

December 2012




[1] Inoue T, Node K. Molecular basis of restenosis and novel issues of drug-eluting stents [J]. Circ J, 2009; 73(4): 615-621.


[2] Ying WZ, Zhang HG, Sanders PW. EGF receptor activity modulates apoptosis induced by inhibition of the proteasome of vascular smooth muscle cells[J]. J Am Soc Nephrol, 2007; 18: 131-142.


[3] Yamanaka Y, Hayashi K, Komurasaki T, et al. EGF family ligand-dependent phenotypic modulation of smooth muscle cells through EGF receptor[J]. Biochem Biophys Res Commun, 2001; 281(2): 373-377.


[4] Cheng Y, Liu X, Yang J, et al. MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation. [J]. Circ Res, 2009; 105(2): 158-166.


[5] Vargas Cruz V, García Ceballos A, Ruiz Hierro C, Moreno Rodríguez MM, et al. Structural alterations of the smooth muscle and of the EGFR expression and C-kit in congenital pyeloureteral stenosis. Relationship with its pathogenesis [J]. Cir Pediatr. 2010 Apr; 23(2): 82-7.

[6] Saltis J, Thomas AC, Agrotis A, et al. Expression of growth factor receptors in arterial smooth muscle cells: Dependency on cell phenotype and serum factors[J]. Atherosclerosis, 1995; 118(1): 77-87.


[7] Zhang H, Chalothorn D, Jackson LF, Lee DC, Faber JE. Transactivation of epidermal growth factor receptor mediates catecholamine-induced growth of vascular smooth muscle[J]. Circ Res. 2004 Nov 12; 95(10): 989-97.


[8] Liao D, Lin PH, Yao Q, et al. Vascular smooth cell proliferation in perfusion culture of porcine carotid arteries[J]. Biochem Biophys Res Commun 2008; 372(4): 668-673.


[9] Horigome M, Kumazaki S, Hattori N, et al. Noninvasive evaluation of coronary endothelial function following sirolimus-eluting stent implantation by using positron emission tomography[J]. Cardiology, 2009; 114(3): 157-63.


[10] Ferguson KM. A Structure-based view of epidermal growth factor receptor regulation[J]. Annu Rev Biophys, 2008; 37: 353-373. Polito L, Bortolotti M, Farini V, et al. Saporin induces multiple death pathways in lymphoma cells with different intensity and timing as compared to ricin[J]. The International Journal of Biochemistry & Cell Biology, 2009; 41(5): 1055-1061.