triple negative breast cancer tnbc cell lines  (ATCC)

Journal: OncoTargets and Therapy

Article Title: Targeting the Inactive Conformation of the Epidermal Growth Factor Receptor Identifies EG31: A Novel Small Molecule Inhibitor Effective Against Normal and 5-Fluorouracil-Resistant Triple Negative Breast Cancer Cells

doi: 10.2147/OTT.S512184

Figure Lengend Snippet: Effect of EG31 in TNBC cells. ( a ) GI 50 values for cell proliferation in EG31-treated MDA-MB-231 and Hs578T cells. ( b ) Effect of EG31 at different doses on the survival of HBL-100 noncancerous breast cells. GraphPad Prism version 6.0 was utilized to examine the mean ± SD values obtained from the MTT assay, which evaluated cell viability and proliferation. ( c ) The proportion of EGFR-positive cells in MDA-MB-231 and Hc578T cells was assessed via flow cytometry. Treatment with EG31 led to a decrease in the EGFR-positive population in both MDA-MB-231 and Hc578T cells. The values presented represent the mean ± standard deviation (SD) from three independent experiments. ( d ) The results of the Annexin V assay demonstrate that EG31 administration induces both early- and late-phase apoptotic cells in MDA-MB-231 and Hs578T cells. Experiments were conducted in triplicate, and the mean ± SD findings are shown. * represents statistical significance; P -value <0.05 compared to respective controls.

Article Snippet: Triple-negative breast cancer (TNBC) cell lines (eg, MDA-MB-231, Hs578T) and non-cancer normal breast cell line (HBL-100) were obtained from ATCC, Rockville, MD, USA, and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: MTT Assay, Flow Cytometry, Standard Deviation, Annexin V Assay

Journal: OncoTargets and Therapy

Article Title: Targeting the Inactive Conformation of the Epidermal Growth Factor Receptor Identifies EG31: A Novel Small Molecule Inhibitor Effective Against Normal and 5-Fluorouracil-Resistant Triple Negative Breast Cancer Cells

doi: 10.2147/OTT.S512184

Figure Lengend Snippet: Antiproliferation in normal and 5-FU resistant TNBC cells. ( a ) Effect of 5-FU in normal MDA-MB-231 cells and 5-FU resistant MDA-MB-231/5-FU R cells in controlling cell proliferation. ( b ) Efficacy of EG31 to control the cell proliferation in the 5-FU resistant MDA-MB-231/5-FU R cells. Cell proliferation was assessed using the MTT assay, and results were analyzed using GraphPad Prism version 6.0 software to determine GI 50 concentration.

Article Snippet: Triple-negative breast cancer (TNBC) cell lines (eg, MDA-MB-231, Hs578T) and non-cancer normal breast cell line (HBL-100) were obtained from ATCC, Rockville, MD, USA, and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: Control, MTT Assay, Software, Concentration Assay

Journal: Cancers

Article Title: Zebrafish Xenografts Unveil Sensitivity to Olaparib beyond BRCA Status

doi: 10.3390/cancers12071769

Figure Lengend Snippet: Zebrafish xenografts of triple negative breast cancer (TNBC) reveal different responses to olaparib and ionizing radiation (IR) independently of BRCA status. TNBC cell lines (Hs578T BRCAwt , MDA-MB-469 BRCAwt and Hcc1937 BRCA1mut ) were fluorescently labeled with CM-DiI (not shown) and injected into the perivitelline space (PVS) of 2dpf zebrafish embryos. At 24 hpi, zebrafish xenografts were screened and randomly distributed into the different experimental conditions: control, olaparib, IR, IR + olaparib. Xenografts were treated for 4 consecutive days and fixed at 5dpi. Representative confocal images of xenografts labelled with anti-human-leukocyte antigen-major histocompatibility complex (HLA-MHC)-class I (in red) ( A,E,I ) and anti-activated caspase 3 (in green) ( A’ – L ). Nuclei were stained with DAPI (in blue) ( A – L ). Mitotic index ( M) , nuclear area size ( N ), cell death-activated caspase3 ( O ) and average tumor size (number of human DAPI cells) ( P ) were analyzed by confocal microscopy and quantified. The % of activated caspase 3 cells and tumor size were normalized to respective controls to compare between different xenografts in different conditions. All images are anterior to the left, posterior to right, dorsal up, and ventral down (as depicted in the scheme on top left). The dashed line delineates the tumor area. Scale bar: 50 µm. Results are from three independent experiments and expressed as mean ± SEM, each dot represents one xenograft. The total number of xenografts analyzed is indicated in the images. Statistical results: ns > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Triple negative breast cancer cell lines Hs578T and MDA-MB-468, originally from American Type Culture Collection, were authenticated through short tandem repeat profiling karyotyping isoenzyme analysis.

Techniques: Labeling, Injection, Control, Immunopeptidomics, Staining, Confocal Microscopy

Journal: Cancers

Article Title: Zebrafish Xenografts Unveil Sensitivity to Olaparib beyond BRCA Status

doi: 10.3390/cancers12071769

Figure Lengend Snippet: Olaparib and IR can have modulating effects on the tumor vascular network. All cell lines were fluorescently labeled with CM-DiI (in red) and injected in the perivitelline space (PVS) of 2dpf Tg(fli1:eGFP) zebrafish embryos. At 24 hpi, zebrafish xenografts were screened and randomly distributed into the different experimental conditions: control, olaparib, IR, IR + olaparib. Xenografts were treated for 4 consecutive days, fixed at 5dpi and imaged by confocal microscopy for analysis of vessel density ( A – T ). The quantification of vessel density is represented by percentage of tumor area occupied by vessels (in green) ( U ). To analyze vessel functionality, TNBC cell line Hs578T was fluorescently labeled with CellTracker™ Deep Red Dye (in red-false color) and injected in the PVS of 2dpf Tg(gata1:RFP;fli1:eGFP). Hs578T xenografts were screened and randomly distributed in the different experimental conditions: control, olaparib, IR and IR + olaparib. Representative images of 5dpi Hs578T xenografts ( V , V’ ). The presence of erythrocytes (in red) inside the tumor-associated vessels (in green) was scored as: absence of erythrocytes = no perfusion ( W , W’ ) or presence of erythrocytes = with perfusion ( X , X’ ) and quantified ( Z ). All images are anterior to the left, posterior to right, dorsal up, and ventral down (as depicted in the scheme on top left). The dashed lines delineate the tumor area. White arrowheads indicate erythrocytes inside the vasculature. Scale bar: 50 µm. Results are from three independent experiments and expressed as mean ± SEM, each dot represents one xenograft. The total number of xenografts analyzed is indicated in the images. Statistical results: ns > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Triple negative breast cancer cell lines Hs578T and MDA-MB-468, originally from American Type Culture Collection, were authenticated through short tandem repeat profiling karyotyping isoenzyme analysis.

Techniques: Labeling, Injection, Control, Confocal Microscopy

Journal: Cancers

Article Title: Zebrafish Xenografts Unveil Sensitivity to Olaparib beyond BRCA Status

doi: 10.3390/cancers12071769

Figure Lengend Snippet: Metastatic potential of TNBC cell lines is modulated by olaparib and IR. TNBC (Hs578T BRCAwt , MDA-MB-469 BRCAwt and Hcc1937 BRCA1mut ) xenografts were generated as previously described and at 24 hpi, were screened and randomly distributed into the different experimental conditions: control, olaparib, IR, IR + olaparib. At 5dpi and 4 days post-treatment, xenografts were fixed to analyze the presence of micrometastasis in the caudal hematopoietic tissue (CHT) ( A ). Representative image of a micrometastasis in the CHT is labelled with anti-human-HLA–MHC-class (in red) for human cell identification. The dashed white line represents the micrometastasis area, scale bar: 50 µm ( A ). The metastatic potential was quantified as the percentage of xenografts that present micrometastasis in the CHT at 5dpi ( B ). Results are from three independent experiments and expressed as mean ± SEM. The total number of xenografts analyzed is indicated below the chart. Statistical results: ns > 0.05, * p ≤ 0.05.

Article Snippet: Triple negative breast cancer cell lines Hs578T and MDA-MB-468, originally from American Type Culture Collection, were authenticated through short tandem repeat profiling karyotyping isoenzyme analysis.

Techniques: Generated, Control