Downregulation of p21(WAF1/CIP1) and estrogen receptor α in MCF-7 cells by antisense oligonucleotides containing locked nucleic acid (LNA)
Downregulation of p21(WAF1/CIP1) and estrogen receptor α in MCF-7 cells by antisense oligonucleotides containing locked nucleic acid (LNA)
复制标题
DOI：
10.1089/1545457041526281
复制
发表时间：
2004-01-01
期刊：
影响因子：
--
通讯作者：
Lykkesfeldt, AE
中科院分区：
文献类型：
--
作者：
Jepsen, JS;Pfundheller, HM;Lykkesfeldt, AE
锁核酸（locked nucleic acid，LNA）是一种与互补RNA有很高亲和力的核酸类似物，是一种很有前途的反义核酸化合物。发现含有LNA的寡核苷酸的细胞内定位和定量摄取与硫代磷酸寡核苷酸（PS AON）的细胞内定位和定量摄取相当。将含LNA的寡核苷酸的反义效率与靶向人乳腺癌细胞系MCF-7中的两种内源性蛋白质（即，细胞周期蛋白依赖性激酶抑制剂p21（（wAF 1/CIP1））和雌激素受体α（ER α））表达的标准PS AON系统地比较。对于两种靶蛋白的下调，最有效的设计是用末端含有LNA单体和PS连接的DNA单体的中心缺口（所谓的LNA缺口聚体）的寡核苷酸实现的。这样的LNA gapmers比PS AON引起靶蛋白更有效的下调，而完全修饰的LNA AON或LNA mixmers（散布的LNA核苷酸）是无活性的。
Locked nucleic acid (LNA) is a nucleic acid analog with very high affinity to complementary RNA and a promising compound in the field of antisense research. The intracellular localization and quantitative uptake of oligonucleotides containing LNA were found to be equivalent to those of phosphorothioate oligonucleotides (PS AONs). The antisense efficiency of LNA-containing oligonucleotides was systematically compared with standard PS AONs targeting expression of two endogenous proteins in the human breast cancer cell line MCF-7, namely, the cyclin-dependent kinase inhibitor p21((wAF1/CIP1)) and the estrogen receptor alpha (ERalpha). For downregulation of both target proteins, the most efficient design was achieved with oligonucleotides containing LNA monomers in the extremities and a central gap of PS-linked DNA monomers, so called LNA gapmers. Such LNA gapmers caused more potent downregulation of the targeted proteins than PS AONs, whereas fully modified LNA AONs or LNA mixmers (LNA nucleotides interspersed) were inactive.