Tan Xing Fei, Teo Wei Xuan, Yip George W
Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
Methods Mol Biol. 2019;1974:21-30. doi: 10.1007/978-1-4939-9220-1_2.
Discovery and development of gene targets for cancer therapeutics are lengthy and highly costly processes. Identification and evaluation of candidate gene targets are of fundamental importance. RNA interference allows candidate genes to be specifically and effectively knocked down in cancer cells. This tool can be easily incorporated into a loss-of-function approach in the initial evaluation of candidate gene targets for cancer treatment prior to moving on to animal studies and clinical trials. This chapter describes a relatively simple and straightforward protocol that makes use of small interfering RNA to achieve knockdown of the candidate gene target and to evaluate the resultant effects on four aspects of cancer cell behavior: migration, invasion, proliferation, and adhesion.
癌症治疗基因靶点的发现与开发是漫长且成本高昂的过程。候选基因靶点的识别与评估至关重要。RNA干扰可使候选基因在癌细胞中被特异性且有效地敲低。在进入动物研究和临床试验之前，该工具可轻松纳入功能丧失方法，用于癌症治疗候选基因靶点的初步评估。本章介绍了一种相对简单直接的方案，该方案利用小干扰RNA实现候选基因靶点的敲低，并评估其对癌细胞行为四个方面的影响：迁移、侵袭、增殖和黏附。
