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Topics {✒️}

hanne roberg-larsen egfr/akt/srebp-1/ldlr-dependent pathway 8p11-p12-amplified breast carcinomas lesley-ann martin pi3k/akt/mtor pathway pi3k/akt/mtor axis [41 low-density lipoprotein receptor article download pdf silac-based quantitative proteomics 8p11-p12 chromosomal region estrogen receptor signaling estrogen receptor-positive long-term estrogen deprivation sep-pak c18 cartridges kaplan–meier plots revealing kaplan-meier plots revealing lc-ms/ms analysis 3-hydroxy-3-methylglutaryl-coa reductase hr + /her2- breast cancer phenol red-free rpmi early-stage breast cancer humanht-12_v4 expression beadchips 27-hydroxycholesterol promotes cell-autonomous long-term oestrogen-deprivation discover protein-dna interactions pure anti-estrogen fulvestrant er ligand-binding domain thousand cell-scale quantification side chain-hydroxylated oxysterols endogenous e-regulated gene full access enhanced estrogen receptor dcc dextran-coated charcoal adipocyte-secreted endocrine factors functional er-transcription axis lbd ligand-binding domain poor long-term outcome individual cholesterol-biosynthesis enzymes patient-level meta-analysis akt-srebp-1-dependent ere-linked luciferase reporter endogenous e-regulated genes lc-ms/ms wild-type er-mediated transcription estrogen deprivation support brb-arraytools data archive proteome-wide protein quantification early breast cancer full-scan spectrum

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         headline:Cholesterol biosynthesis pathway as a novel mechanism of resistance to estrogen deprivation in estrogen receptor-positive breast cancer
         description:Therapies targeting estrogenic stimulation in estrogen receptor-positive (ER+) breast cancer (BC) reduce mortality, but resistance remains a major clinical problem. Molecular studies have shown few high-frequency mutations to be associated with endocrine resistance. In contrast, expression profiling of primary ER+ BC samples has identified several promising signatures/networks for targeting. To identify common adaptive mechanisms associated with resistance to aromatase inhibitors (AIs), we assessed changes in global gene expression during adaptation to long-term estrogen deprivation (LTED) in a panel of ER+ BC cell lines cultured in 2D on plastic (MCF7, T47D, HCC1428, SUM44 and ZR75.1) or in 3D on collagen (MCF7) to model the stromal compartment. Furthermore, dimethyl labelling followed by LC-MS/MS was used to assess global changes in protein abundance. The role of target genes/proteins on proliferation, ER-mediated transcription and recruitment of ER to target gene promoters was analysed. The cholesterol biosynthesis pathway was the common upregulated pathway in the ER+ LTED but not the ER– LTED cell lines, suggesting a potential mechanism dependent on continued ER expression. Targeting the individual genes of the cholesterol biosynthesis pathway with siRNAs caused a 30–50 % drop in proliferation. Further analysis showed increased expression of 25-hydroxycholesterol (HC) in the MCF7 LTED cells. Exogenous 25-HC or 27-HC increased ER-mediated transcription and expression of the endogenous estrogen-regulated gene TFF1 in ER+ LTED cells but not in the ER– LTED cells. Additionally, recruitment of the ER and CREB-binding protein (CBP) to the TFF1 and GREB1 promoters was increased upon treatment with 25-HC and 27-HC. In-silico analysis of two independent studies of primary ER+ BC patients treated with neoadjuvant AIs showed that increased expression of MSMO1, EBP, LBR and SQLE enzymes, required for cholesterol synthesis and increased in our in-vitro models, was significantly associated with poor response to endocrine therapy. Taken together, these data provide support for the role of cholesterol biosynthesis enzymes and the cholesterol metabolites, 25-HC and 27-HC, in a novel mechanism of resistance to endocrine therapy in ER+ BC that has potential as a therapeutic target.
         datePublished:2016-06-01T00:00:00Z
         dateModified:2016-06-01T00:00:00Z
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         keywords:
            Breast cancer
            Estrogen receptor
            Cholesterol biosynthesis
            Oxysterol
            Transcriptomics
            Proteomics
            Cancer Research
            Oncology
            Surgical Oncology
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      headline:Cholesterol biosynthesis pathway as a novel mechanism of resistance to estrogen deprivation in estrogen receptor-positive breast cancer
      description:Therapies targeting estrogenic stimulation in estrogen receptor-positive (ER+) breast cancer (BC) reduce mortality, but resistance remains a major clinical problem. Molecular studies have shown few high-frequency mutations to be associated with endocrine resistance. In contrast, expression profiling of primary ER+ BC samples has identified several promising signatures/networks for targeting. To identify common adaptive mechanisms associated with resistance to aromatase inhibitors (AIs), we assessed changes in global gene expression during adaptation to long-term estrogen deprivation (LTED) in a panel of ER+ BC cell lines cultured in 2D on plastic (MCF7, T47D, HCC1428, SUM44 and ZR75.1) or in 3D on collagen (MCF7) to model the stromal compartment. Furthermore, dimethyl labelling followed by LC-MS/MS was used to assess global changes in protein abundance. The role of target genes/proteins on proliferation, ER-mediated transcription and recruitment of ER to target gene promoters was analysed. The cholesterol biosynthesis pathway was the common upregulated pathway in the ER+ LTED but not the ER– LTED cell lines, suggesting a potential mechanism dependent on continued ER expression. Targeting the individual genes of the cholesterol biosynthesis pathway with siRNAs caused a 30–50 % drop in proliferation. Further analysis showed increased expression of 25-hydroxycholesterol (HC) in the MCF7 LTED cells. Exogenous 25-HC or 27-HC increased ER-mediated transcription and expression of the endogenous estrogen-regulated gene TFF1 in ER+ LTED cells but not in the ER– LTED cells. Additionally, recruitment of the ER and CREB-binding protein (CBP) to the TFF1 and GREB1 promoters was increased upon treatment with 25-HC and 27-HC. In-silico analysis of two independent studies of primary ER+ BC patients treated with neoadjuvant AIs showed that increased expression of MSMO1, EBP, LBR and SQLE enzymes, required for cholesterol synthesis and increased in our in-vitro models, was significantly associated with poor response to endocrine therapy. Taken together, these data provide support for the role of cholesterol biosynthesis enzymes and the cholesterol metabolites, 25-HC and 27-HC, in a novel mechanism of resistance to endocrine therapy in ER+ BC that has potential as a therapeutic target.
      datePublished:2016-06-01T00:00:00Z
      dateModified:2016-06-01T00:00:00Z
      pageStart:1
      pageEnd:14
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      sameAs:https://doi.org/10.1186/s13058-016-0713-5
      keywords:
         Breast cancer
         Estrogen receptor
         Cholesterol biosynthesis
         Oxysterol
         Transcriptomics
         Proteomics
         Cancer Research
         Oncology
         Surgical Oncology
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      name:The Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK
      name:The Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK
      name:Academic Department of Biochemistry, Royal Marsden Hospital, London, UK
      name:Proteomics Core Facility, Institute of Cancer Research, London, UK
      name:Proteomics Core Facility, Institute of Cancer Research, London, UK
      name:The Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK
      name:Academic Department of Biochemistry, Royal Marsden Hospital, London, UK
      name:The Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK

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