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MiR-200c induction in Met-1 mammary carcionma cells

ABSTRACT: miR-200c induction in Met-1 mammary carcionma cells

PROVIDER: PRJNA635432 | ENA |

REPOSITORIES: ENA

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miR-200c induction in Met-1 mammary carcionma cells

Project description:We performed RNA-seqeuncing analysis on mouse Met-1 cells that express miR-200c in a doxycycline (DOX)-inducible manner from the miR-200c-TripZ system.

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miR-200c induction in BT549 TNBC cells

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Essential role of miR-200c in regulating self-renewal of breast cancer stem cells initiating from the counter parts of mammary epithelium

Project description:To explore the roles of essential miRNAs in regulating self-renewal of breast cancer stem cells (BCSCs), which initiate from mammary epithelial stem cells (MaSCs). CD44+CD24-/low cells and MUC1-ESA+ cells were isolated by fluorescence-activated cell sorting (FACS) from breast cancer cell line MCF-7 and normal mammary epithelial cell line MCF-10A, and were verified as BCSCs and MaSCs by clonogenic assay and multipotential differentiation experiment in 2-dimensional (2-D) and 3-D cultures, respectively. Using microarray containing oligonucleotides corresponding to 509 miRNAs from human, mouse, and rat genomes. We obtained candidate miRNAs in regulating breast tumorigenesis. One representative miRNA (miR-200c) was proved to regulate stemness of BCSCs and MaSCs in vitro and in vivo by miR-200c agomir transfection. We validated that miR-200c negatively regulated PDCD10, an apoptosis regulator, in BCSCs and MaSCs.

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Project description:To investigate the changes of gene expression in mouse mammary gland after knockout of miR-200c/141 in mammary luminal cells. Luminal cells from CKO (K8-CreERT2;miR-200c/141 fl/fl) and Ctrl (K8-CreERT2;miR-200c/141 fl/+) mice were FACS-isolated for RNA-seq analysis.

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Effect of miR-200c-3p overexpression on primary human macrophages

Project description:Macrophages constitute a major part of the tumor-infiltrating immune cells and within the tumor microenvironment acquire an alternatively activated, tumor-supporting phenotype. Factors released by tumor cells are crucial for the recruitment of tumor-associated macrophages. In the present project, we aimed to understand the role of miR-200c in the interplay between tumor cells and macrophages. To this end, we employed a coculture system of MCF7 breast tumor cells and primary human macrophages and observed a substantial transfer of miR-200c from apoptotic tumor cells to macrophages, which required intact CD36 receptor in macrophages. We further comprehensively determined miR-200c targets in macrophages by mRNA-sequencing and found numerous migration-associated mRNAs to be downregulated by miR-200c. Consequently, miR-200c attenuated macrophage infiltration into 3-dimensional tumor spheroids. The miR-200c-mediated reduction of infiltration further correlated well with a miR-200c migration signature comprised of four miR-200c-repressed targets (PPM1F, RAB11FIB2, RDX, MSN).

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Expression data from differentiating ES cells expressing miR-200c and miR-141

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TALEN-based knockout of mir-141 and mir-200c in SK-BR-3 cells

Project description:We used transcription activator-like effector nucleases (TALENs) to generate knockout cells for two related microRNAs (miRNAs), mir-141 and mir-200c, which belong to the deeply conserved mir-200 family. By carrying out deep sequencing, we identified the target genes of each miRNA. Interestingly, miR-141 and miR-200c, despite their overall similarity, suppressed largely non-overlapping groups of targets. Analysis of global mRNA level change in mir-141 and mir-200c knockout compared to wild type cells

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