论文部分内容阅读
目的研究硫代寡核苷酸的亲和性及其在内皮细胞株的吸收与亚细胞分布。方法 利用电泳迁移分析法检测硫代寡核苷酸的亲和性;将同位素标记的硫代寡核苷酸与ECV304细胞直接孵化,24 F后测定内皮细胞的吸收率及亚细胞分布。结果3条硫代寡核苷酸(T21GTa、T14Gta及T15Gta)的亲和性(Kd值)分别为2.3×10-9、3.8×10-9及1.5×10-8(M).细胞吸收率分别为11.9％、10.8％、11.1％,核沉淀放射活性分别为77.25％、76.75％、69.32％。结论3条硫代寡核苷酸能够与内皮细胞TF基因SSRE的3个Egr-1/Spl位点结合,具有较好的透膜性、耐酶性及稳定性,能够透过内皮细胞膜和核膜,抵达核内靶基因部位。
Objective To study the affinity of thio-oligonucleotide and its absorption and subcellular distribution in endothelial cell lines. Methods The electrophoretic mobility shift assay (MTA) was used to detect the affinity of thio-oligonucleotide. The isotope-labeled thio-oligonucleotide was incubated directly with ECV304 cells. The absorbance and subcellular distribution of endothelial cells were measured after 24-F incubation. Results The affinity (Kd) values of the three thio oligonucleotides (T21GTa, T14Gta and T15Gta) were 2.3 × 10-9, 3.8 × 10-9 and 1.5 × 10-8 (M), respectively. Respectively 11.9%, 10.8% and 11.1%, and the nuclear sediment radioactivity was 77.25%, 76.75% and 69.32% respectively. CONCLUSION: Three thio-oligonucleotides can bind to the three Egr-1 / Spl sites of the TF gene of endothelial cells and have good permeability, enzyme resistance and stability. They can permeate the endothelial cell membrane and nucleus Membrane, reaching the nuclear target site.