Project description:Analysis of reciprocal modulation of Runx2 and E2 signaling in BCA. Our objective was to investigate whether the interaction between estrogen and Runx2 signaling in breast cancer (BCa) could help refine an estrogen-responsive gene signature with improved prognostic value. MCF7/Rx2dox BCa cells conditionally expressing Runx2 upon doxycycline treatment were treated with estradiol and/or doxycycline to induce Runx2, and global gene expression was profiled to define genes regulated by estradiol, Runx2, or both.
Project description:Analysis of reciprocal modulation of Runx2 and E2 signaling in BCA. Our objective was to investigate whether the interaction between estrogen and Runx2 signaling in breast cancer (BCa) could help refine an estrogen-responsive gene signature with improved prognostic value.
Project description:Catechol-O-methyl transferase (COMT) is involved in detoxification of catechol estrogens, playing cancer-protective role in cells producing or utilizing estrogen. Moreover, COMT suppressed migration potential of breast cancer cells. To delineate COMT role in metastasis of estrogen receptor dependent BC, we investigated the effect of COMT overexpression on invasion, transcriptome, proteome and interactome of MCF7 cells, a luminal A breast cancer model, stably transduced with lentiviral vector carrying COMT gene (MCF7-COMT). This PRIDE project includes quantitative analysis results for the total proteome LC-DIA-MS/MS experiment evaluating COMT overexpression in MCF7 breast cancer cell line, and results of pulldown analysis of COMT-interacting proteins in MCF7 cells.
Project description:Endocrine therapy resistance is a clinical problem for the management of estrogen receptor (ER)-positive breast cancer. The elucidation of factors that modulate ER signaling will provide useful information for understanding the pathophysiology of ER-positive and endocrine-resistant breast cancers. In the screen of estrogen-inducible lncRNAs transcribed from ER alpha-associated active promoters/enhancers, a novel estrogen-inducible lncRNA BNAT1 (breast cancer natural antisense transcript 1), which is transcribed from the proximal promoter region of COL18A1, was identified in ER+ MCF7 cells. siRNA-mediated BNAT1 silencing significantly inhibited the in vitro and in vivo growth of tamoxifen-resistant MCF7 cells.To examine the role of long non-coding RNA BNAT1 (alias gene name COL18AS was used in this dataset) in ER+ breast cancer, MCF7 cells were treated with siRNAs targeting BNAT1 (siCOL18AS, or siBNAT1) or control siRNA (siCtrl, or siControl). The microarray study showed that BNAT1 was closely associated with estrogen signaling pathway.
Project description:Previous studies have identified an interaction between RUNX1 and estrogen receptor alpha and its potential role in estrogen signaling in breast cancer. To determine the transcriptomic actions of RUNX1, we knocked down its expression by using siRNAs, both in the absence and presence of estradiol (E2).
Project description:Estrogen signaling plays important roles in diverse physiological and pathophysiological processes. However, the relationship between estrogen signaling and epigenetic regulation is not fully understood. Here, we explored the effect of estrogen signaling on the expression of Ten-Eleven Translocation (TET) family genes and DNA hydroxylmethylation in estrogen receptor alpha positive (ERα+) breast cancer cells. By analyzing the RNA-seq data, we identified TET2 as an estradiol (E2)-responsive gene in ERα+ MCF7 cells. RT-qPCR and Western blot analyses confirmed that both the mRNA and protein levels of TET2 gene were upregulated in MCF7 cells by E2 treatment. ChIP-seq and qPCR analyses showed that the enrichment of ERα and H3K27ac on the upstream regulatory regions of TET2 gene was increased in MCF7 cells upon E2 treatment. Moreover, E2 treatment also led to a significant increase in the global 5-hydroxymethylcytosine (5hmC) level, while knockout of TET2 abolished such E2-induced 5hmC increase. Conversely, treatment with ICI 182780, a potent and selective estrogen receptor degrader (SERD), inhibited TET2 gene expression and down-regulated the 5hmC level in MCF7 cells. Taken together, our study identified an ERα/TET2/5hmC epigenetic pathway, which may participate in the estrogen-associated physiological and pathophysiological processes.
Project description:The genes for transcription factor RUNX1 and its binding partner CBFb have been reported to be mutated in human breast cancer. We show for the first time that loss of Runx1 in a genetically engineered mouse model results in accelerated disease onset and tumour development aligning RUNX1 as a tumour suppressor in breast cancer. Combined deletion of Runx1 and the related family member Runx2 resulted in mammary epithelial cells becoming exquisitely sensitive to WNT-driven transformation with the emergence of multiple tumours early in life. Clonogenic assays indicated that Runx1 ablation induced a stem cell like phenotype in mammary epithelial cells whilst transcriptome analysis demonstrated activation of multiple oncogenic pathways, especially when Runx2 was co-deleted. While loss of Runx2 itself did not result in tumour promotion we observed dramatic effects when both Runx2 and Runx1 were deleted. Significantly, altered Runx expression in the mammary epithelium also drove profound alterations in the tumour microenvironment impacting the immune landscape revealing extrinsic effects of Runx gene deletion in mammary tissue. Tissue, mouse model: Cell preparations from 9-week mammary glands were from female mice of the following genotypes: 1) BLG-Cre;Catnbwt/lox(ex3) 2) BLG-Cre;Catnbwt/lox(ex3);Runx1fl/fl 3) BLG-Cre;Catnbwt/lox(ex3);Runx2fl/fl 4) BLG-Cre;Catnbwt/lox(ex3); Runx1fl/fl;Runx2fl/fl
