The effect of transcription factor SP1 decoy oligodeoxynucleotides on expression of α Gal in SV-40-PED cells

Objective:To investigate the role of transcription factor SP1 decoy oligodeoxynucleotides(ODN) on expression of α Gal in SV-40-PED cells.Methods:Immortalized porcine aortic endothelial cells of the PED line were cultured and transfected with α1,3galactosyltransferase(α1,3GT) specific decoy ODN.Cells transfected with mismatch ODN was used as negative controls.Twenty-six hours later the cells were collected.The expression of α Gal was determined with fluorescence microscope and Western blot.The expression of α1,3GT mRNA was examined by RT-PCR.Results:Fluorescence microscopy observed the decreased fluorescence of α Gal after decoy ODN transfection.Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was(48.2±0.9).It is 52.6% of the mock group(P0.05).The expression of α1,3GT mRNA in the PED cells transfected with decoy ODN was(isoform 1,0.42±0.20;isoform 2,0.27±0.12) lower in comparison with the mock group(isoform 1,(0.72)±0.17;isoform 2,0.50±0.19;both P0.05).The expression of α1,3GT in the mismatch group was not different from those in the mock group(P0.05).Conclusion:α1,3GT gene reduce actually occurs following transfection of decoy ODN.Porcine endothelial cells can be the targets of decoy ODN.