Pitts A E, Corey D R
Howard Hughes Medical Institute, Departments of Pharmacology and Biochemistry, University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Boulevard, Dallas, TX 75235-9050, USA.
Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11549-54. doi: 10.1073/pnas.95.20.11549.
Telomerase, a ribonucleoprotein up-regulated in many types of cancers, possesses an RNA template necessary to bind and extend telomere ends. The intrinsic accessibility of telomerase to incoming nucleic acids makes the RNA template an ideal target for inhibition by oligonucleotides. We report here that 2'-O-methyl-RNA (2'-O-meRNA), an oligonucleotide chemistry known to exert sequence-specific effects in cell culture and animals, inhibits telomerase with potencies superior to those possessed by analogous peptide nucleic acids (PNAs). Potent inhibition relative to PNAs is surprising, because the binding affinity of 2'-O-meRNAs for complementary RNA is low relative to analogous PNAs. A 2'-O-meRNA oligomer with terminal phosphorothioate substitutions inhibits telomerase sequence-selectively within human-tumor-derived DU145 cells when delivered with cationic lipids. In contrast to the ability of 2'-O-meRNA oligomers to inhibit telomerase, the binding of a 2'-O-meRNA to an inverted repeat within plasmid DNA was not detectable, whereas binding of PNA was efficient, suggesting that the relative accessibility of the telomerase RNA template is essential for inhibition by 2'-O-meRNA. Inhibition of telomerase by 2'-O-meRNA will facilitate probing the link between telomerase activity and sustained cell proliferation and may provide a basis for the development of chemopreventive and chemotherapeutic agents.
端粒酶是一种在多种癌症中上调的核糖核蛋白,它拥有结合并延长端粒末端所需的RNA模板。端粒酶对进入的核酸具有内在的可及性,这使得RNA模板成为寡核苷酸抑制的理想靶点。我们在此报告,2'-O-甲基RNA(2'-O-meRNA)是一种在细胞培养和动物中能发挥序列特异性作用的寡核苷酸化学物质,它抑制端粒酶的效力优于类似的肽核酸(PNA)。相对于PNA的强效抑制作用令人惊讶,因为2'-O-meRNA与互补RNA的结合亲和力相对于类似的PNA较低。当与阳离子脂质一起递送时,具有末端硫代磷酸酯取代的2'-O-meRNA寡聚物在人肿瘤来源的DU145细胞内对端粒酶进行序列选择性抑制。与2'-O-meRNA寡聚物抑制端粒酶的能力相反,未检测到2'-O-meRNA与质粒DNA内反向重复序列的结合,而PNA的结合则很有效,这表明端粒酶RNA模板的相对可及性对于2'-O-meRNA的抑制作用至关重要。2'-O-meRNA对端粒酶的抑制将有助于探究端粒酶活性与细胞持续增殖之间的联系,并可能为化学预防和化学治疗药物的开发提供基础。