Antiproliferative activity of G-quartet-forming oligonucleotides with backbone and sugar modifications
Antiproliferative activity of G-quartet-forming oligonucleotides with backbone and sugar modifications
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DOI：
10.1021/bi0119520
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发表时间：
2002-03-19
期刊：
影响因子：
2.9
通讯作者：
Miller, DM
中科院分区：
文献类型：
--
作者：
Dapic, V;Bates, PJ;Miller, DM
基于寡核苷酸的疗法在癌症、病毒和心血管疾病治疗中具有相当大的潜力。然而，越来越清楚的是，寡核苷酸的生物学效应不仅仅归因于与核酸的预期序列特异性相互作用。由于寡核苷酸的聚阴离子特性或特定的二级结构，寡核苷酸还能够与许多细胞蛋白质相互作用。我们检查了一系列富含鸟苷的寡核苷酸的抗增殖活性、蛋白质结合和 G 四联体形成，这些寡核苷酸是 GRO29A 的类似物，GRO29A 是一种 G 四联体形成、生长抑制性寡核苷酸，其作用我们之前已经描述过 [Bates P. J.、Kahlon, J. B.、Thomas, S. D.、Trent, J. O. 和 Miller, D. M. (1999) J.生物。化学。 274、26369-26377]。 GRO29A 类似物包括硫代磷酸酯 (PS29A)、2'-O-甲基 RNA (MR29A) 和混合 DNA/2'-O-甲基 RNA (MRdG29A) 寡核苷酸。我们通过紫外光谱证明所有修饰的类似物形成稳定的结构，这与 G-四联体的形成一致。我们发现硫代磷酸酯和混合DNA/2'-O-甲基类似物能够显着抑制多种肿瘤细胞系的增殖，而2'-O-甲基RNA则没有显着效果。与原始寡核苷酸 GRO29A 类似，生长抑制寡核苷酸能够与人端粒序列寡核苷酸竞争结合特定的细胞蛋白质。活性较低的 MR29A 不会显着竞争该蛋白质。根据寡核苷酸结构的分子建模，MR29A的失活很可能是由于该寡核苷酸形成的四链体的凹槽结构的差异所致。有趣的是，所有 GRO29A 类似物，包括未修饰的 DNA 磷酸二酯寡核苷酸，在含血清培养基存在下均能显着抵抗核酸酶降解，表明二级结构在生物稳定性中发挥着重要作用。这些寡核苷酸显着的稳定性和强大的抗增殖活性证实了它们作为治疗剂的潜力。
Oligonucleotide-based therapies have considerable potential in cancer, viral, and cardiovascular disease therapies. However, it is becoming clear that the biological effects of oligonucleotides are not solely due to the intended sequence-specific interactions with nucleic acids. Oligonucleotides are also capable of interacting with numerous cellular proteins owing to their polyanionic character or specific secondary structure. We have examined the antiproliferative activity, protein binding, and G-quartet formation of a series of guanosine-rich oligonucleotides, which are analogues of GRO29A, a G-quartet forming, growth-inhibitory oligonucleotide, whose effects we have previously described [Bates P. J., Kahlon, J. B., Thomas, S. D., Trent, J. O., and Miller, D. M. (1999) J. Biol. Chem. 274, 26369-26377]. The GRO29A analogues include phosphorothioate (PS29A), 2'-O-methyl RNA (MR29A), and mixed DNA/ 2'-O-methyl RNA (MRdG29A) oligonucleotides. We demonstrate by UV spectroscopy that all of the modified analogues form stable structures, which are consistent with G-quartet formation. We find that the phosphorothioate and mixed DNA/2'-O-methyl analogues are able to significantly inhibit proliferation in a number of tumor cell lines, while the 2'-O-methyl RNA has no significant effects. Similar to the original oligonucleotide, GRO29A, the growth inhibitory oligonucleotides were able to compete with the human telomere sequence oligonucleotide for binding to a specific cellular protein. The less active MR29A does not compete significantly for this protein. On the basis of molecular modeling of the oligonucleotide structures, it is likely that the inactivity of MR29A is due to the differences in the groove structure of the quadruplex formed by this oligonucleotide. Interestingly, all GRO29A analogues, including an unmodified DNA phosphodiester oligonucleotide, are remarkably resistant to nuclease degradation in the presence of serum-containing medium, indicating that secondary structure plays an important role in biological stability. The remarkable stability and strong antiproliferative activity of these oligonucleotides confirm their potential as therapeutic agents.