Project description:To screen potential target genes of SPIN1 in breast cancer, human gene expression microarray was performed to identify differentially expressed mRNAs in SPIN1 siRNA or negative control transfected MCF-7/ADM cells. After transfection with SPIN1 siRNA, 4409 genes were differentially expressed by >2-fold in MCF-7/ADM cells compared with negative control-transfected cells, of which 2413 were significantly downregulated and the other 1996 were upregulated. Differentially expressed genes were submitted to GO and KEGG pathway analysis.
Project description:Transcriptional profiling of breast cancer cell line MCF-7 stably transfected with pcDNA3.1-Gli1 comparing control ones transfected with CON
Project description:Overexpression of the AP-2γ transcription factor in breast tumours has been identified as an independent predictor of poor outcome and failure of hormone therapy, even in ER positive, ErbB2 negative tumours; markers of a more favourable prognosis. To understand further the role of AP-2γ in breast carcinoma, we have used an RNA interference and gene expression profiling strategy using the MCF-7 cell line as a model for ER positive, ErbB2 negative tumours with AP-2γ overexpression. Gene expression changes between control and silenced cells implicate AP-2γ in the control of cell cycle progression and developmental signalling. Experiment Overall Design: We compared the expression profiles of MCF-7 cells separately transfected with three independent AP-2γ targeting sequences with those from control cells treated with transfection reagent alone or a non-silencing control siRNA on Affymetrix arrays; each condition was examined in triplicate.
Project description:Copy number loss of AKTIP gene is frequently observed in breast cancer particularly luminal breast cancer. To determine the alterations in transcriptome upon AKTIP loss in luminal breast cancer cell line MCF7 and triple negative breast cancer cell line SKBR3, RNAseq of cells transfected with AKTIP siRNA was performed.
Project description:PRMT5 Co-IP mass spectrometry: Lysates of MCF-7 RB1-knockout cells were pulled down with a PRMT5 antibody or IgG control. The pulldowns were subjected to mass spectrometry analysis.
QEX2_981005, QEX2_981006, QEX2_981007: DMSO, IgG
QEX2_981008, QEX2_981009, QEX2_981010: DMSO, PRMT5 antibody
QEX2_981011, QEX2_981012, QEX2_981013, GSK, IgG
QEX2_981014, QEX2_981015, QEX2_981016, GSK, PRMT5 antibody
SDMA PTM analysis: MCF-7 RB1-knockout cells were transfected with a PRMT5 siRNA or a control siRNA. Lysates were collected three days after transfection and subjected to mass spectrometry analysis.
LUM1_1000791, LUM1_1000792, LUM1_1000793: control SiRNA
LUM1_1000794, LUM1_1000795, LUM1_1000796: PRMT5 siRNA
Project description:PRMT5 Co-IP mass spectrometry: Lysates of MCF-7 RB1-knockout cells were pulled down with a PRMT5 antibody or IgG control. The pulldowns were subjected to mass spectrometry analysis. QEX2_981005, QEX2_981006, QEX2_981007: DMSO, IgG QEX2_981008, QEX2_981009, QEX2_981010: DMSO, PRMT5 antibody QEX2_981011, QEX2_981012, QEX2_981013, GSK, IgG QEX2_981014, QEX2_981015, QEX2_981016, GSK, PRMT5 antibody SDMA PTM analysis: MCF-7 RB1-knockout cells were transfected with a PRMT5 siRNA or a control siRNA. Lysates were collected three days after transfection and subjected to mass spectrometry analysis. LUM1_1000791, LUM1_1000792, LUM1_1000793: control SiRNA LUM1_1000794, LUM1_1000795, LUM1_1000796: PRMT5 siRNA [
Project description:The transcription factor Snail has been proposed to mediate epithelial-to-mesenchymal transition (EMT) and confer mesenchymal invasive phenotype to epithelial cancer cells To analyze the molecular effects of ectopic Snail expression on an epithelial breast cancer cell line, gene expression profiles of MCF-7 cells transfected to overexpress Snail-6SA variant (MCF-7-Snail) and MCF-7 cells transfected with control plasmid (MCF-7-control) were compared. Development of the cell lines has been previously reported by Zhou et al. (PMID: 15448698).
Project description:MCF-7 is an estrogen receptor-positive breast cancer cell line. This experiment is designed to study (1) the effect of estradiol (E2) exposure and (2) lysine methyltransferase 2B (KMT2B) knockdown in MCF-7 cells. Cells were grown for 72 hours prior to treatment with vehicle or 10 nM E2 for 4 and 24 hours. Additionally, to assess the effect of KMT2B knockdown, MCF-7 cells were transfected with KMT2B targeting siRNA or scrambled control siRNA in the absence or presence of E2. RNA were isolated using Trizol and hybridized to Affymetrix GeneChip Human Genome U133 Plus 2.0 array.
Project description:HSD3B1 (3beta-hydroxysteroid dehydrogenase type 1) plays a vital role in steroidogenesis. Transcription profiling analysis was performed in MDA-MD-231 breast cancer cells transfected with negative control siRNA or siRNAs against HSD3B1.
Project description:The telomeric amplicon at 8p12 is common in ER+ breast cancers. Array-CGH and expression analyses of 1172 tumors revealed ZNF703/Zeppo1 was the single gene within the minimal amplicon and was amplified predominantly in the Luminal B subtype. Amplification was shown to correlate with increased gene and protein expression and was associated with a distinct expression signature and poor outcome. In the luminal MCF-7 cell line manipulation of ZNF703 expression altered transcription of genes also present within the primary tumor signature, including TGFBR2 (whose promoter was bound by ZNF703). Overexpression of ZNF703 rendered MCF-7 cells insensitive to TGFβ-induced suppression of mammosphere formation. Forced overexpression of ZNF703 in normal human breast epithelial cells enhanced the frequency of in vitro colony-forming cells from luminal progenitors. Together these data strongly point to ZNF703/Zeppo1 as a novel oncogene in Luminal B breast cancer. MCF-7 breast cancer cell line was infected with ZNF703 overexpression (ZNF703) or control (HIV) virus and following GFP sorting of infected cells, were transfected with control siRNA (siC) or siRNA against endogenous ZNF703 (siZNF), resulting in four different conditions: siC_HIV, siC_ZNF, siZNF_HIV and siZNF-ZNF. RNA for each condition was harvested from triplicate plates.