Involvement of post-transcriptional regulation of FOXO1 by HuR in 5-FU-induced apoptosis in breast cancer cells

The post-transcriptional control of specific mRNAs is a widespread mechanism of gene regulation, which contributes to numerous biological processes in a number of cell types. The Forkhead box O (FoxO) transcription factor FOXO1 is an important tumor suppressor involved in apoptosis, the cell cycle, DNA damage repair and oxidative stress. Bioinformatic prediction identified that the 3' untranslated region (UTR) of FOXO1 is enriched with binding motifs for the human ELAV/Hu protein (HuR), indicating that FOXO1 is a potential target of HuR. Luciferase reporter assays demonstrate that HuR specifically regulates FOXO1 expression through AU-rich elements (AREs) within the FOXO1 3' UTR. Immunoprecipitation studies confirmed that HuR associates with FOXO1 mRNA in MDA-MB-231 breast cancer cells and that HuR upregulates FOXO1 mRNA levels through increased mRNA stability. Using a HuR loss- and gain-of-function approach, we revealed that FOXO1 expression was correspondingly decreased or increased in MDA-MB-231 cells. Functional assays demonstrated that HuR and FOXO1 expression levels were markedly enhanced upon 5-fluorouracil (5-FU) stimulation in MDA-MB-231 cells. Knockdown of HuR apparently abrogated 5-FU-induced apoptosis detected by caspase-3 activities. Furthermore, in HuR knockdown cells, additional overexpression of FOXO1 moderately recovered 5-FU-induced apoptosis, which verified that HuR-modulated apoptosis upon 5-FU treatment was partially mediated by its post-transcriptional regulation of FOXO1. Therefore, modulating FOXO1 expression has been suggested to lead to the development of new therapeutic treatments for certain types of cancer.