Development of a high-content screening method to evaluate the cellular population-based biological activity of new siRNA delivering reagent  

高内涵筛选用于评价新型siRNA运载试剂的细胞群体生物学活性(英文)

作　　者：李雅婷[1] 郑宜[1] 潘德林[1] 徐波[1] 武芸[1] 杨振军[1] 张礼和[1] 

机构地区：[1]北京大学医学部 天然药物及仿生药物国家重点实验室,北京100191

出　　处：《Journal of Chinese Pharmaceutical Sciences》2013年第6期467-474,共8页中国药学（英文版）

基　　金：Ministry of Science and Technology of China(Grant No.2012CB720604);NSFC(Grant No.20932001)

摘　　要：Normally, cellular responses to modified siRNAs or new siRNA delivery systems have been studied in group cell behavior by PCR, western blotting and fluorescence microscopy. In this study, we present a novel high-content screening （HCS） strategy to evaluate a novel delivery system （named CLD） of siRNA therapeutics, with which both the content of intracellular siRNAs and changes in protein expressing levels have been quantified in group cells and cellular population. We also observed that with the better cell uptake, CLD provided siRNA therapeutics （siBraf） better antitumor capability. This novel strategy was proved to be with efficiency, accuracy and high competency to adherent cell lines, thus making siRNA research more simplified.修饰siRNAs和新型siRNA运载系统对细胞的作用通常通过PCR、Western Blotting和荧光显微镜来进行细胞整体特征的研究。本文报道了基于高内涵、用于siRNA治疗的一种新型运载系统的新策略(CLD),该策略从细胞整体和细胞群体水平综合考虑siRNA的转染效率及对蛋白表达水平的影响。我们证明CLD能达到更佳的细胞摄取,使siBraf达到更好的抗肿瘤活性。该方法具有高效、准确和适用于多个贴壁细胞系的优点,能简化siRNA的研究过程。

关 键 词：High-content screening siRNA delivery system Gene knock-down Cellular population ERK signaling pathway 

分 类 号：R944[医药卫生—药剂学]