Project description:Overexpression in IL6 in MCF10A vs. WT, MCF10A.ErbB2* with/without shRNA targeting STAT3 Overexpression in IL6 in MCF10A vs. WT, MCF10A.ErbB* with/without shSTAT3, duplicates, two STAT3 shRNAs
Project description:C/EBPbeta-2 results in EMT and ErbB indpendence this project investigated the gene changes in related genes upon C/EBPbeta-2 overexpression in MCF10A cells. We used microarray analysis to detail the global gene expression mediated by C/EBPbeta-2 and identified changes in known EMT genes, however, known ErbB related genes were not altered. MCF10A with or without C/EBPbeta-2 were compared.
Project description:To knock down SLC3A2, MCF10A cells were either transfected with control siRNAs, or siRNAs specifically targetting SLC3A2. Samples were either untreated or treated with TGFb. Ribosome profiling and RNA sequencing was performed on these samples. MCF10A cells with and without SLC3A2 overexpression construct were either untreated or treated with TGFb. Ribosome profiling was performed on these samples.
Project description:To identify gene expression changes associated with overexpression of miR-105 or MYC in MCF10A non-cancerous human mammary epithelial cells, we analyzed RNA isolated from engineered MCF10A cell lines that stably express empty vector, GFP, miR-105, or MYC by RNA-seq. Gene expression in cells overexpressing miR-105 or MYC was compared to cells expressing the empty vector or GFP, both of which served as controls in this experiment.
Project description:PHF8 exerts distinct functions in different types of cancer. However, the mechanisms underlying its specific functions in each case remain obscure. To establish whether overexpression of PHF8 regulates the TGF-β induced the epithelial-mesenchymal transition (EMT), we treated MCF10A-Mock (control) and MCF10A-PHF8wt (overexpressing wild-type PHF8) cells with TGF-β1 for 0, 24, 48 and 72 hours and performed RNA-seq in biological duplicates. Our data indicated that EMT gene signatures were significantly enriched in MCF10A-PHF8 cells with TGF-β1 treatment at all time points, strongly indicating that PHF8 overexpression induces a sustained EMT signaling program.
Project description:PHF8 exerts distinct functions in different types of cancer. However, the mechanisms underlying its specific functions in each case remain obscure. To establish whether overexpression of PHF8 regulates the TGF-β induced the epithelial-mesenchymal transition (EMT), we treated MCF10A-Mock (control) and MCF10A-PHF8wt (overexpressing wild-type PHF8) cells with TGF-β1 for 0, 24, 48 and 72 hours and performed RNA-seq in biological duplicates. Our data indicated that EMT gene signatures were significantly enriched in MCF10A-PHF8 cells with TGF-β1 treatment at all time points, strongly indicating that PHF8 overexpression induces a sustained EMT signaling program. mRNA profiles of MCF10A-Mock (control) and MCF10A-PHF8 with TGF-β1 treatment for 0, 24, 48 and 72 hours were generated by RNA-seq, in duplicate, using HiSeq2500 instrument.
Project description:Excessive expression of subunit 1 of GIRK1 in ER+ breast tumors is associated with reduced survival times, and increased lymph node metastasis in patients. To investigate possible tumor-initiating properties, benign MCF10A and malign MCF7 mammary epithelial cells overexpressing GIRK1 were engineered, neoplasia associated vital parameters and resting potentials were measured and compared to controls. Presence of GIRK1 resulted in resting potentials negative to controls. Upon GIRK1 overexpression, several cellular pathways were sizably regulated towards pro-tumorigenic action as revealed by comparison of transcriptomes of MCF10AGIRK1 with control (MCF10AeGFP). According to transcriptome analysis, cellular migration was promoted while wound healing and extracellular matrix interactions were impaired. Vital parameters in MCF7 cells were affected akin the benign MCF10A lines, although to a lesser extent. Thus, GIRK1 overexpression will aid in classification of ER+ breast tumors and bears the potential to open a new window for the therapy of this detrimental disease.