Project description:We report mRNA profiles of human breast cancer cell lines, MCF7 parental, and MCF7-derived tamoxifen resistant cell lines MCF7-TR1 and MCF7-TR2.
Project description:Quiescent breast cancer stem cells (Q-BCSCs) can switch to an activated state (A-BCSCs). We established a dual-reporter system for stemness and quiescence (NANOG-EGFP + H2B-mCherry pulse-chase) to isolate A-BCSCs and Q-BCSCs from MDA-MB-231 cells. Strand-specific poly(A) RNA-seq was performed to compare A-BCSCs vs Q-BCSCs (n=3 per group). In a second dataset, ENO1 silencing (siENO1) was compared with a non-targeting control (siNC) in unsorted MDA-MB-231 cells (n=2 per group). Processed data are provided as separate experiment-specific gene-level matrices rather than a single unified matrix because different Ensembl gene annotation releases were used for quantification in the two experiments. For the A-BCSCs vs Q-BCSCs dataset, counts and FPKM matrices together with a matching gene annotation table are provided. For the siENO1 vs siNC dataset, counts and FPKM matrices together with a matching gene annotation table are provided. Previous reports have identified NANOG as a key stemness factor in tumor cells. Based on this, we established a stable fluorescent reporter system in MDA-MB-231 cells via lentiviral transduction, in which EGFP is driven by the NANOG promoter and expressed upon NANOG activation. To validate whether this system faithfully reflects cellular stemness, NANOG-EGFP positive and negative cells were sorted by FACS and subsequently analyzed by RNA-seq. Our results showed that the NANOG-EGFP positive population was enriched for stemness-related gene pathways, confirming that this reporter system can effectively indicate tumor cell stemness
Project description:Immuno-precipitation followed by MS was performed using the RIME protocol in this study. Estrogen Receptor (ER) and Progestorone Receptor (PR) were targetted using antibodies. The experiments were performed in a quantitative manner using SILAC labelling. Cells grown in complete serum (estrogenic) conditions were compared against an cells in similar complete media, however supplemented with Progesterone (PG) or R5020 for 4 hours. Experiments were performed in MCF7 and T47D cell lines