Project description:Gene expression analysis for structural components in Rb-deficient and RB-proficient ErbB2-overexpressing immortalized breast cell line using 3-D cell culture model to analyze structural and organizational changes of epithelial cells. Cells were cultured in a reconstituted membrane for 7 days. Cells were extracted with the use RNA-STAT-60 method.
Project description:Gene expression analysis for structural components in Rb-deficient and RB-proficient ErbB2-overexpressing immortalized breast cell line using 3-D cell culture model to analyze structural and organizational changes of epithelial cells.
Project description:Gene expression profiling of ErbB2-engineered MCF10A and WT cells in 2D and 3D culture Gene expression profiling of ErbB2-engineered MCF10A and WT cells, 2D and 3D culture, 5 or 6 replicates
Project description:ErbB2 activation of MCF10A cells gives rise to a multiacinar phenotype that is incompletely penetrant. To identify candidate upstream regulators of the phenotype, an MCF10A-5E clone was subcloned to express chimeric receptors of human EGFR and rat Erbb2, and this B2B1 subclone was stochastically profiled (PMID: 20228812) after 24 hours of 3D culture with or without synthetic dimerization.
Project description:Approximately 30% of TNBCs exhibit loss of function of the RB tumor suppressor. The study used RNA sequencing to profile RB-proficient and RB-deficient TNBC cases that were defined based on immunostaining for RB and p16ink4a. The analyses revealed that RB-deficient TNBC cases express elevated levels of DNA replication and mitotic genes that could serve as the basis for increased sensitivity to drugs targeting cell cycle checkpoints.
Project description:Most BRCA1-deficient BLBCs carry a dysfunctional INK4-RB pathway. Thus, we have created genetically engineered mice with Brca1 loss and deletion of p16INK4A, or separately p18INK4C, to model the deficient INK4-RB signaling in human BLBC. By using these mutant mice and human BRCA1 deficient and proficient breast cancer tissues and cells, we tested if there exists a druggable target in BRCA1 deficient breast cancers.
Project description:This project uses an MCF10A clone (B2B1) expressing chimeric synthetic receptors for ErbB1 (EGFR) and ErbB2 (HER2) along with BirA*-fused inducible alleles of CSE1L or NUP37. Cells were cultured with reconstituted basement membrane, BirA* alleles induced with doxycycline, ErbB receptors dimerized with a synthetic small molecule, and biotinylation was performed for 24 hours. Lysates underwent a two-step biotin affinity purification before gel electrophoresis and mass spectrometry on selected gel fragments.
Project description:Expression data from ERBB2 over-expression and EGF stimulation in MCF10A cells The cells were transduced with retroviruses encoding vector control (pBabe) or pBabe-ErbB2. After infection cells were switched to assay medium supplemented with only 2% horse serum and no EGF. For EGF stimulation, cells transduced with empty vector were treated with EGF at 50 ng/ml for 2 hours before harvesting.
