Perez J R, Li Y, Stein C A, Majumder S, van Oorschot A, Narayanan R
Division of Oncology, Roche Research Center, Hoffmann-La Roche, Inc., Nutley, NJ 07110.
Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5957-61. doi: 10.1073/pnas.91.13.5957.
Modified analogues of antisense oligodeoxynucleotides (ODNs), particularly phosphorothioates ([S]ODNs), have been extensively used to inhibit gene expression. The potential sequence specificity of antisense oligomers makes them attractive as molecular drugs for human diseases. The use of antisense [S]ODNs to inhibit gene expression has been complicated by frequent nonspecific effects. In this study we show in diverse cell types that [S]ODNs, independent of their base sequence, mediated the induction of an Sp1 nuclear transcription factor. The [S]ODN-mediated Sp1 induction was rapid and was associated with elevated levels of Sp1 protein. This induction was dependent on NF-kappa B activity, since inhibition of NF-kappa B activity abolished the [S]ODN-induced Sp1 activity. [S]ODN-induced Sp1 activity was seen in mouse spleen cells following in vivo administration. Sp1 activity induced by [S]ODNs required the tyrosine kinase pathway and did not have transactivating potential. These results may help to explain some of the non-specific effects often seen with [S]ODNs.
反义寡脱氧核苷酸（ODNs）的修饰类似物，特别是硫代磷酸酯（[S]ODNs），已被广泛用于抑制基因表达。反义寡聚物潜在的序列特异性使其作为治疗人类疾病的分子药物具有吸引力。使用反义[S]ODNs抑制基因表达因频繁的非特异性效应而变得复杂。在本研究中，我们在多种细胞类型中表明，[S]ODNs与其碱基序列无关，介导了Sp1核转录因子的诱导。[S]ODN介导的Sp1诱导迅速，且与Sp1蛋白水平升高有关。这种诱导依赖于NF-κB活性，因为抑制NF-κB活性可消除[S]ODN诱导的Sp1活性。体内给药后，在小鼠脾细胞中可观察到[S]ODN诱导的Sp1活性。[S]ODNs诱导的Sp1活性需要酪氨酸激酶途径，且不具有反式激活潜能。这些结果可能有助于解释[S]ODNs经常出现的一些非特异性效应。
