ABSTRACT
In the published version of this article, the images for cytoplasmic and nuclear FGF7 in MDA-MB-231 cells were duplicated and mistaken for total FGF7 in SKBR-3 and MDA-MB-231 cells.
ABSTRACT
Vascular endothelial growth factor (VEGF) has a central role in breast cancer development and progression, but the mechanisms that control its expression are poorly understood. Breast cancer tissue microarrays revealed an inverse correlation between the Forkhead transcription factor Forkhead box class O (FOXO)3a and VEGF expression. Using the lapatinib-sensitive breast cancer cell lines BT474 and SKBR3 as model systems, we tested the possibility that VEGF expression is negatively regulated by FOXO3a. Lapatinib treatment of BT474 or SKBR3 cells resulted in nuclear translocation and activation of FOXO3a, followed by a reduction in VEGF expression. Transient transfection and inducible expression experiments showed that FOXO3a represses the proximal VEGF promoter, whereas another Forkhead member, FOXM1, induces VEGF expression. Chromatin immunoprecipitation and oligonucleotide pull-down assays showed that both FOXO3a and FOXM1 bind a consensus Forkhead response element (FHRE) in the VEGF promoter. Upon lapatinib stimulation, activated FOXO3a displaces FOXM1 bound to the FHRE before recruiting histone deacetylase 2 (HDAC2) to the promoter, leading to decreased histones H3 and H4 acetylation, and concomitant transcriptional inhibition of VEGF. These results show that FOXO3a-dependent repression of target genes in breast cancer cells, such as VEGF, involves competitive displacement of DNA-bound FOXM1 and active recruitment of transcriptional repressor complexes.
Subject(s)
Breast Neoplasms/metabolism , Forkhead Transcription Factors/metabolism , Vascular Endothelial Growth Factor A/metabolism , Cell Line, Tumor , Female , Forkhead Box Protein M1 , Forkhead Box Protein O3 , Gene Expression Regulation, Neoplastic , Histone Deacetylase 2/metabolism , Humans , Lapatinib , Quinazolines/pharmacologyABSTRACT
Amino acid uptake into leaf fragments of Pisum sativum depended on metabolism. Glycine uptake was optimal at 30 C and could be supported by respiration and by photosynthesis. Based on studies with an electron flow cofactor, inhibitors, and uncouplers, the energy source for glycine uptake was apparently ATP.The energy-dependent transport of glycine was mediated by a carrier that had a broad specificity for neutral and positively charged l-amino acids. It readily translocated 15 such l-amino acids into the cells, but had a very low affinity for l-aspartate, l-glutamate, d-amino acids, and alpha-aminoisobutyrate. The Ki for competitive inhibition of glycine uptake by another amino acid was equal to the Km for the uptake of that competing species.
