ArrayExpress Uploader0mouseOnmouse mammary tumor derived cell line 8534UntreatedOffmouse mammary tumor derived cell line 8534doxycyclineOnmouse mammary tumor derived cell line 8542UntreatedOffmouse mammary tumor derived cell line 8542doxycycline6707arrayexpress_sid6NOTCH activation has been recently implicated in human basal-like breast cancers associated with a poor prognosis. To address the role of...E-GEOD-34146/profile/8773/arrayexpressuploader34adenocarcinomasbasalbreastcelldiseaseembryonic stem cellgenomekeratinleftmammary tumornotchstem cellDec.12, 2014Falsetranscriptional-profiling-of-mmtv-ttatop-icn1-tumoE-GEOD-34146_A-AFFY-45Transcriptional profiling of MMTV-tTA/TOP-ICN1 tumor derived cell lines comparing untreated (ICN1-On) cells to 24-hour doxcycline treated (ICN1-Off) samplesNov.12, 2014NOTCH activation has been recently implicated in human basal-like breast cancers associated with a poor prognosis. To address the role of Notch1 in mammary transformation and mammary tumor initiating cell activity, we developed a doxycycline-regulated model of Notch1-mediated mammary transformation. These mice develop mammary adenocarcinomas that express cytokeratin (CK) 8/18 and contain rare cells that also express keratin 14. In vivo limiting dilution analyses reveal that these mammary tumors exhibit functional heterogeneity and harbor a rare (1/2978) mammary tumor initiating cell population. Using this dox-regulated Notch1 mammary tumor model, we demonstrate that Notch1 inhibition results in mammary tumor regression in vivo and prevents disease recurrence in 4 of 6 tumors tested. Consistent with the in vivo data, Notch1 inhibition reduces mammary tumorsphere forming activity in vitro. Using doxycycline-responsive tumor derived cell lines, we also identify the embryonic stem cell transcription factor Nanog as a novel Notch1-regulated gene in mammospheres. These data indicate that Notch1 contributes to mammary tumor initiating activity and raises the possibility that NOTCH therapeutics may have efficacy in human basal-like breast cancers associated with NOTCH activation. Primary mammary tumors were isolated from two different MMTV-tTA/TOP-ICN1 transgenic mice, minced, enzymatically digested and converted to culture. To identify changes in gene expression in response to ICN1 suppression, tumor-derived cell lines 8534 and 8542 were left untreated (8534-Untreated; 8542-Untreated) or treated with 2ug/ml doxycycline for 24 hours (8534-Dox; 8542-Dox). Cells were collected by scraping and total RNA was isolated, followed by real-time PCR validation of NOTCH1 target gene modulation. RNA samples were further hybridized to Affymetrix mouse genome 430A2.0 arrays.http://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-34146http://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-34146/samples/