Journal: Breast Cancer Research : BCR

Article Title: Phosphorylated and sumoylation-deficient progesterone receptors drive proliferative gene signatures during breast cancer progression

doi: 10.1186/bcr3211

Figure Lengend Snippet: The SUMO-deficient PR gene expression signature is associated with HER2-positive human breast tumors and predicts reduced patient survival . ( A ) Normalized gene expression levels (for genes in our LD KR > WT gene signature) are presented for each tumor in the patient cohort , organized by ERBB2 status. ( B ) Gene expression levels were measured by RT-qPCR for CHN2 and RGS2 (both upregulated by SUMO-deficient PR, and members of the LD KR > WT gene signature) and the control gene ACOT6 (equally upregulated by both WT and KR receptors) in BT-474 human breast cancer cells. Cells were pre-treated with MEK kinase inhibitor U0126 prior to progestin or antiprogestin co-treatment. Protein levels were evaluated by western blotting for total PR, PR Ser294 phosphorylation, total ERK1/2, and ERK1/2 phosphorylation. ( C ) Kaplan-Meier survival curve for distant metastasis free survival for patients whose tumors expressed the combined T47D metagenes (WT or KR, -/+R5020) relative to patient tumors lacking these metagenes. Patient samples include untreated and tamoxifen-treated ER-positive tumors from the Loi et al. dataset . ( D ) Survival curves as in part C for patients whose tumors expressed the combined T47D metagenes (KR -R5020, or KR +R5020) relative to patient tumors lacking these metagenes. [See also Additional files and ]. ER, estrogen receptor; KR, K388R PR-B mutant; LD, ligand dependent; SUMO, small ubiquitin-like modifier; WT, wild type.

Article Snippet: BT-474 cells (ATCC, Manassas, VA, USA) were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Grand Island, NY, USA, catalog #11875) supplemented with 10% FBS, 1% penicillin/streptomycin.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Mutagenesis

Journal: bioRxiv

Article Title: Elucidating Fibroblast Growth Factor-induced kinome dynamics using targeted mass spectrometry and dynamic modeling

doi: 10.1101/2023.01.27.525819

Figure Lengend Snippet: MCF-7, BT-474, and EFM-192a cells were stimulated with 50ng/mL of either FGF2 or FGF4 supplemented with 5µg/mL of heparin. After 0, 30, and 60 minutes, the cells were harvested and subjected to measurement using the targeted kinome assay. The heatmap shows the quantified activation-determining phosphorylated sites on the kinases (biological triplicates). Only significantly changing values are shown (ANOVA p < 0.05).

Article Snippet: MCF-7 (ATCC), BT-474 (ATCC), and EFM-192a (DSMZ) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS (Sigma) and 2mM glutamine.

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Elucidating Fibroblast Growth Factor-induced kinome dynamics using targeted mass spectrometry and dynamic modeling

doi: 10.1101/2023.01.27.525819

Figure Lengend Snippet: FGFR expression levels were quantified using qPCR in MCF-7, BT-474, and EFM-192a cells using FGFR subtype-specific primers (triplicate measurements) ( Supplementary table 5 ). Beta-actin and Glyceraldehyde-3-phosphate dehydrogenase was quantified to enable normalization across cell types. Reported values are the quantitation cycles (Cq) that negatively correlates with the RNA expression levels.

Article Snippet: MCF-7 (ATCC), BT-474 (ATCC), and EFM-192a (DSMZ) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS (Sigma) and 2mM glutamine.

Techniques: Expressing, Quantitation Assay, RNA Expression

Journal: International Journal of Nanomedicine

Article Title: The Combined Effect of Nanobubble-IR783-HPPH-Affibody Complex and Laser on HER2-Positive Breast Cancer

doi: 10.2147/IJN.S387409

Figure Lengend Snippet: The targeting ability of the NIHA complex. The CLSM images of BT-474 cells incubated with the NIHA complex (up) and the nanobubble-Dil-HPPH complex (down). Bar, 50 μm.

Article Snippet: The BT-474 cell line and the MDA-MB-231 cell line were purchased from the American Type Culture Collection (USA).

Techniques: Incubation

Journal: International Journal of Nanomedicine

Article Title: The Combined Effect of Nanobubble-IR783-HPPH-Affibody Complex and Laser on HER2-Positive Breast Cancer

doi: 10.2147/IJN.S387409

Figure Lengend Snippet: ( A ) Cell viability of BT-474 cells detected via CCK-8. ( B ) The apoptotic ratio of BT-474 cells detected via Annexin-V flow cytometry. ** P < 0.01 compared with the NIHA-laser group.

Article Snippet: The BT-474 cell line and the MDA-MB-231 cell line were purchased from the American Type Culture Collection (USA).

Techniques: CCK-8 Assay, Flow Cytometry

Journal: International Journal of Nanomedicine

Article Title: The Combined Effect of Nanobubble-IR783-HPPH-Affibody Complex and Laser on HER2-Positive Breast Cancer

doi: 10.2147/IJN.S387409

Figure Lengend Snippet: CLSM images of BT-474 cells in the NIHA-laser, NIHA, laser, and the control groups. The nuclei of the apoptosis cells were stained by using PI (depicted as red fluorescence). Bar, 30 μm.

Article Snippet: The BT-474 cell line and the MDA-MB-231 cell line were purchased from the American Type Culture Collection (USA).

Techniques: Staining, Fluorescence

Journal: Nature Communications

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

doi: 10.1038/s41467-023-35856-y

Figure Lengend Snippet: MCF-7 and BT-474 cells were cultured on soft (0.5 kPa) or stiff (9 kPa) supports for 2 weeks before subsequent analysis. a Docetaxel (Doc) induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. The proportions of Annexin V + cells are shown. b Quantitation of apoptotic cells (Annexin V + ) in MCF-7 and BT-474 cells in (a) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. c MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated after 24 h by CCK8 assays ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. d Western blots for cleaved/uncleaved caspase-3 and PARP in docetaxel-treated MCF-7 and BT-474 cells cultured on soft or stiff supports. Representative images from 3 biologically independent experiments are shown. e The percentage of CD44 high CD24 low cells in MCF-7 and BT-474 cells cultured on soft or stiff supports were determined by flow cytometry. f Quantitation of CD44 high CD24 low cells in (e) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. For b-d and f , source data are provided as a Source Data file.

Article Snippet: MCF-7 cells (cat#HTB-22) and BT-474 cells (cat#HTB-20) cells were obtained from American Type Culture Collection (ATCC) and cultured according to standard protocols.

Techniques: Cell Culture, Flow Cytometry, Quantitation Assay, Western Blot

Journal: Nature Communications

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

doi: 10.1038/s41467-023-35856-y

Figure Lengend Snippet: MCF-7 and BT-474 cells were cultured on soft or stiff supports for 2 weeks before subsequent analysis. In some experiments, cells were pre-transduced with TAZ shRNA (shTAZ-1 or shTAZ-2) or GFP shRNA (shGFP). a The expression of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff supports was determined by western blot ( n = 3 biologically independent experiments). b Representative immunofluorescence staining of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff substrates. The insert represents a magnification from the indicated area (white square). Scale bars, 10 μm. c Proportion of cancer cells exhibiting mainly nuclear TAZ localization (N > C), diffuse distribution of TAZ in nucleus and cytoplasm (N = C), or preferential cytoplasmic TAZ (N < C or undetectable). 20 cells were assessed per group, experiments were independently repeated 4 times. Mean ± SD, **** p < 0.0001 by two-tailed chi-square test. d Docetaxel induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. See Supplementary Fig. for quantification. e MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated by CCK8 assay ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided one-way ANOVA with Tukey test. f Representative images of CD44 high CD24 low cells in MCF-7 and BT-474 cells were detected by flow cytometry. See Supplementary Fig. for quantification. g Western blots of TAZ, SOX2, and OCT4 in MCF-7 and BT-474 cells cultured on soft or stiff supports ( n = 3 biologically independent experiments). For data presented in the same figure panel, the samples were derived from the same experiment and blots were processed in parallel. For a , c , e and g , source data are provided as a Source Data file.

Article Snippet: MCF-7 cells (cat#HTB-22) and BT-474 cells (cat#HTB-20) cells were obtained from American Type Culture Collection (ATCC) and cultured according to standard protocols.

Techniques: Cell Culture, Transduction, shRNA, Expressing, Western Blot, Immunofluorescence, Staining, Two Tailed Test, Flow Cytometry, CCK-8 Assay, Derivative Assay

Journal: Nature Communications

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

doi: 10.1038/s41467-023-35856-y

Figure Lengend Snippet: a – d TAZ and NANOG were knockout or overexpressed by lentiviral transfection in MCF-7 and BT-474 cells. NANOG, TAZ, SOX2 or/and OCT4 were overexpressed in TAZ or NANOG knockout cells. Transfected cells were cultured on stiff hydrogels for 2 weeks. OE, overexpression. a (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five biologically independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0003, ** p = 0.0039, ns=0.9899; for BT-474 cells, *** p = 0.0008, ns=0.9982. b Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgTAZ + Vector group were shown in the figure. c (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0004, ** p = 0.0033, ns=0.9400; for BT-474 cells, *** p = 0.0002, ** p = 0.0013, ns=0.9982. d Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgNANOG + Vector group were shown in the figure. For panels (a– d) , data are mean ± SD; p values were calculated by two-sided one-way ANOVA with Tukey test. e , f MCF-7 cells were cultured on soft or stiff hydrogels for 2 weeks. Then ChIP was performed using cell lysates with indicated antibodies. e The precipitated DNA using IgG and TAZ antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . f The precipitated DNA using IgG and NANOG antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . e , f Fold enrichment was normalized to IgG-ChIP negative control ( n = 4 biologically independent experiments). Numeric values denote mean ± SD; **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. For a–f , source data are provided as a Source Data file.

Article Snippet: MCF-7 cells (cat#HTB-22) and BT-474 cells (cat#HTB-20) cells were obtained from American Type Culture Collection (ATCC) and cultured according to standard protocols.

Techniques: Knock-Out, Transfection, Cell Culture, Over Expression, Plasmid Preparation, Negative Control

Journal: BMC Cancer

Article Title: Genetic variability in MCF-7 sublines: evidence of rapid genomic and RNA expression profile modifications

doi: 10.1186/1471-2407-3-13

Figure Lengend Snippet: Hierarchical clustering of RNA expression profiles. Panel A clustering analysis of expression profiles of 8 MCF-7 sublines along with those of 19 breast cancer cell lines. Expression profiling was done using home made Nylon arrays comprising 721 cDNAs corresponding to identified genes localized on either chromosome 1q or 17q. Panel B clustering analysis of profiles of 7 MCF-7 sublines and the BT-474 breast cancer cell line. Nylon arrays comprised 1034 genes selected on the basis of their involvement in cancer. Clustering analysis wass done on raw quantification results, which were just subjected to a scaling step but not to ratio calculation. Parameters used in the analysis were Hierarchically Cluster Axes for Genesand Array: clusterand similarity metric correlation centered with average linkage clustering. The dendogram on top of the diagram represents cell lines ordered according to their degree of similarity. Complete datasets can be found at

Article Snippet: Seven MCF-7 variants (MCF-7-MVLN, MCF-7-ATCC, MCF-7-MF, MCF-7-MG, MCF-7-R, MCF-7-R-F3 and MCF-7-R-D4) were analyzed together with BT-474 breast cancer cells.

Techniques: RNA Expression, Expressing