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目的设计合成靶向NF-κB的哑铃形诱骗剂ODN,分析靶向NF-κB的哑铃形诱骗剂对NF-κB转录活性、多发性骨髓瘤细胞8266的生长及其分泌的IL-6的抑制效应。方法采用电泳迁移率变动分析(EMSA)体外检测诱骗剂ODN对NF-κB转录活性的抑制效应。将8266细胞随机分为传代培养的8266细胞组;诱骗剂ODN处理组及脂质体处理组。通过阳离子脂质体以2mg/L、4mg/L、8mg/L不同剂量诱骗剂ODN转染8266细胞。转染后8、12、18h,ELISA法检测8266细胞培养上清中IL-6的表达。用MTT比色检查诱骗剂ODN对IL-6刺激的8266细胞生长的影响。结果硫代磷酸的诱骗剂ODN在体外能有效地抑制NF-κB与其顺式元件的结合;2mg/L、4mg/L、8mg/L等不同浓度的脂质体-ODN复合物对8266细胞表达IL-6的抑制程度不同。脂质体-ODN复合物对8266细胞的生长及IL-6的活性均有抑制作用。结论靶向NF-κB的诱骗剂ODN在体外可抑制NF-κB的转录活性,从而抑制8266细胞的生长,降低瘤细胞中IL-6的表达。
OBJECTIVE: To design a dumbbell decoy ODN targeting to NF-κB and to study the effect of dumbbell-shaped decoy targeting NF-κB on NF-κB transcriptional activity, growth of multiple myeloma cell line 8266 and secretion of IL-6 effect. Methods The inhibitory effect of decoy ODN on the transcriptional activity of NF-κB was examined by electrophoretic mobility shift assay (EMSA) in vitro. The 8266 cells were randomly divided into subcultured 8266 cells, ODN-treated groups and liposome-treated groups. 8266 cells were transfected with different doses of decoy ODN by cationic liposomes at 2mg / L, 4mg / L, 8mg / L. The expression of IL-6 in 8266 cell culture supernatant was detected by ELISA at 8, 12, 18 h after transfection. The effect of decoy ODN on IL-6-stimulated 8266 cell growth was examined by MTT colorimetric assay. Results Phosphorothioate decoy ODN could effectively inhibit the binding of NF-κB to its cis-elements in vitro. The liposome-ODN complexes of 2mg / L, 4mg / L, 8mg / L and so on could express 8266 cells The degree of inhibition of IL-6 is different. Liposome-ODN complex inhibited the growth of 8266 cells and the activity of IL-6. Conclusion ODN targeting NF-κB can inhibit the transcriptional activity of NF-κB in vitro and thus inhibit the growth of 8266 cells and decrease the expression of IL-6 in tumor cells.