E-GEOD-48158 - Identification of miRNA targets in breast cancer cells (Ago2-IP)
Released on 21 October 2013, last updated on 3 June 2014
miRNAs regulate mRNA stability and translation through the action of the RNAi-induced silencing complex. In this study, we systematically identified endogenous miRNA target genes by using AGO2 immunoprecipitation (AGO2-IP) and microarray analyses in two breast cancer cell lines, MCF7 and MDA-MB-231, representing luminal and basal-like breast cancer, respectively. The expression levels of ~70% of the AGO2-IP mRNAs were increased by DROSHA or DICER1 knockdown. In addition, integrated analysis of miRNA expression profiles, mRNA-AGO2 interaction, and the 3'-UTR of mRNAs revealed that >60% of the AGO2-IP mRNAs were putative targets of the fifty most abundantly expressed miRNAs. To identify mRNAs associated with AGO2, cell lysate was precleaned with control IgG and immunoprecipitated with anti-human Ago2 (Clone 2E12-1C9, Abnova). Total RNA from cell lysate or coimmunoprecipitated with AGO2 was extracted with Trizol and subjected to microarray analysis, with three biological repeats for each experimental condition.
transcription profiling by array
Aarti Sethuraman, Chuan H Yang, Jing Sun, Junming Yue, Lawrence M Pfeffer, Meiyun Fan, Raisa Krutilina, Zhaohui Wu
Comprehensive Analysis of MicroRNA (miRNA) Targets in Breast Cancer Cells. Fan M, Krutilina R, Sun J, Sethuraman A, Yang CH, Wu ZH, Yue J, Pfeffer LM. , Europe PMC 23921383