Synthesis of a triple helix-forming phosphorothioate oligodeoxynucleotides and its effects on coagulation activity of tissue factor (TF) and TF gene expression in endothelial cells.
- Author:
Qianning LI
1
;
Dajun YING
;
Guangming DAI
;
Jian ZHENG
Author Information
1. Biomechanical Laboratory, Department of Anatomy, Third Military Medical University, Chongqing 400038.
- Publication Type:Journal Article
- MeSH:
Blood Coagulation;
drug effects;
Cells, Cultured;
Endothelium;
cytology;
drug effects;
Gene Expression;
Humans;
Oligodeoxyribonucleotides;
chemical synthesis;
pharmacology;
RNA, Messenger;
biosynthesis;
Thromboplastin;
biosynthesis;
genetics;
physiology
- From:
Journal of Biomedical Engineering
2003;20(1):71-90
- CountryChina
- Language:Chinese
-
Abstract:
This study sought to synthesize a triple helix-forming phosphorothioate oligodeoxynucleotides (TFO-ps) and assess its effects on coagulation activity of the tissue factor(TF) and TF gene expression in endothelial cells. Experiment antiparallel oligodeoxynucleotides T21GTa sequence was designed and synthesized by phosphoramidite method and decorated with all-PS linkage. The affinity of TFO and TFO-ps was determined by electrophoretic mobility shift assays(EMSA). Cellular uptake of [32]P-labeled TFO-ps and the effect of TFO-ps on TF gene expression and coagulation activity of TF were measured in endothelial cell strain ECV304. The results showed that TFO-ps (T21GTa-ps) formed a triplex binding in antiparallel orientation to the puring-rich target strand-SSRE, with Kd value of 3.6 x 10(-10) M. The uptake rate of TFO-ps (T21GTa-ps) in ECV304 was about 11.65%. This compound mainly localized within the nucleus sediment(77.25%), significantly reduced the average OD of the mRNA expression and protein synthesis of TF gene, and obviously decreased the coagulation activity of TF. In conclusion, TFO-ps (T21GTa-ps) shows obvious anti-coagulation activity and its mechanism involves the inhibition of TF gene expression.
