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EMT induction in MCF7 cells by overexpression of SNAIL transcription factor

ABSTRACT: The human breast cancer cell line MCF7 was trasnduced with SNAIL trasncription factor. In-depth proteomic analysis revealed several important cellular processes regulated, especially related to epigenetic control of cell proliferation and protein expression.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Vitor Faca

LAB HEAD: Vitor Faca

PROVIDER: PXD002449 | Pride | 2016-01-20

REPOSITORIES: Pride

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			Action	DRS
	140203_VF_CP_MCF7_CCTR_S01.mzXML	Mzxml
	140203_VF_CP_MCF7_CCTR_S01.xtan.xml	Xml
	140203_VF_CP_MCF7_CCTR_S02.mzXML	Mzxml
	140203_VF_CP_MCF7_CCTR_S02.xtan.xml	Xml
	140203_VF_CP_MCF7_CCTR_S04.mzXML	Mzxml

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Publications

Proteomic Analysis of Epithelial to Mesenchymal Transition (EMT) Reveals Cross-talk between SNAIL and HDAC1 Proteins in Breast Cancer Cells.

Palma Camila de Souza Cde S Grassi Mariana Lopes ML Thomé Carolina Hassibe CH Ferreira Germano Aguiar GA Albuquerque Daniele D Pinto Mariana Tomazini MT Ferreira Melo Fernanda Ursoli FU Kashima Simone S Covas Dimas Tadeu DT Pitteri Sharon J SJ Faça Vitor M VM

Molecular & cellular proteomics : MCP 20160113 3

Epithelial to mesenchymal transition (EMT)(1) occurs naturally during embryogenesis, tissue repair, cancer progression, and metastasis. EMT induces cellular and microenvironmental changes resulting in loss of epithelial and acquisition of mesenchymal phenotypes, which promotes cellular invasive and migratory capabilities. EMT can be triggered by extracellular factors, including TGF-β, HGF, and EGF. Overexpression of transcription factors, such as SNAIL, SLUG, ZEB1/2, and TWIST1, also induces EMT ...[more]

PMID: 26764010

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Project description:Epithelial-Mesenchymal Transition (EMT) is thought to contribute to cancer metastasis, but its underlying mechanisms are not well understood. To define early steps in this cellular transformation, we analyzed human mammary epithelial cells with tightly regulated expression of Snail-1, a master regulator of EMT. Following Snail-1 induction, epithelial markers were repressed within 6 hours and mesenchymal genes induced at 24 hours. Snail-1 binding to its target promoters was transient (6-48 hours) despite continued protein expression and it was followed by both transient and long-lasting chromatin changes. To generate a potent reversible EMT-inducing stimulus, we created a Snail-1 retroviral expression construct, using a fused estrogen receptor (ER) response element to mediate regulation by exogenous 4-hydroxy-tamoxifen (4-OHT). Since Snail-1 protein stability and nuclear localization are suppressed by GSK3-beta-mediated phosphorylation, we substituted the six targeted amino acids (ER-Snail-1(6SA)), thus conferring constitutive activity to the induced protein (Zhou et al., 2004, Pubmed ID 15448698). Infection of non-transformed, immortalized human mammary epithelial MCF10A cells with ER-Snail-1(6SA), followed by treatment with 4-OHT, triggered morphological and biomarker characteristics of EMT. At 0, 6, 48 and 120 hours after beginning exposure to 4-OHT in ethanol (or, for controls, ethanol only) we ChIPed Snail-1 and 6 histone marks. We perfomed two replicates of each, except we only had one replicate of H3K27Me3 at 6 hours.

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