Brognara Eleonora, Fabbri Enrica, Bianchi Nicoletta, Finotti Alessia, Corradini Roberto, Gambari Roberto
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Methods Mol Biol. 2014;1095:165-76. doi: 10.1007/978-1-62703-703-7_14.
The involvement of microRNAs in human pathologies is a firmly established fact. Accordingly, the pharmacological modulation of their activity appears to be a very appealing issue in the development of new types of drugs (miRNA therapeutics). One of the most interesting issues is the possible development of miRNA therapeutics for development of anti-cancer molecules. In this respect appealing molecules are based on peptide nucleic acids (PNAs), displaying a pseudo-peptide backbone composed of N-(2-aminoethyl)glycine units and found to be excellent candidates for antisense and antigene therapies. The major limit in the use of PNAs for alteration of gene expression is the low uptake by eukaryotic cells. The aim of this chapter is to describe methods for determining the activity of PNAs designed to target oncomiRNAs, using as model system miR-221 and its target p27(Kip1) mRNA. The effects of PNAs targeting miR-221 are here presented discussing data obtained using as model system the human breast cancer cell line MDA-MB-231, in which miR-221 is up-regulated and p27(Kip1) down-regulated.
微小RNA参与人类疾病已成为确凿事实。因此，对其活性进行药理学调控在新型药物（微小RNA疗法）研发中似乎是一个极具吸引力的课题。最有趣的课题之一是开发用于抗癌分子的微小RNA疗法。在这方面，基于肽核酸（PNA）的分子颇具吸引力，其具有由N-（2-氨基乙基）甘氨酸单元组成的假肽骨架，被发现是反义疗法和反基因疗法的极佳候选物。使用PNA改变基因表达的主要限制是真核细胞摄取率低。本章旨在描述以miR-221及其靶标p27（Kip1）mRNA为模型系统，测定靶向致癌微小RNA的PNA活性的方法。本文介绍了靶向miR-221的PNA的作用，并讨论了使用人乳腺癌细胞系MDA-MB-231作为模型系统获得的数据，在该细胞系中miR-221上调而p27（Kip1）下调。
