Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: Therapeutic window analysis of curcumin and gemcitabine in normal (MCF-10A) and cancer cells. ( A ) Concentration-dependent effects of curcumin and gemcitabine on MCF-10A cell viability following 48 h exposure. Curcumin exhibited moderate cytotoxicity, whereas gemcitabine showed a stronger dose–response effect. The IC 50 reference line (red dashed) and the therapeutic threshold (70%, green dotted) are indicated. The shaded green area represents the proposed therapeutic window. ( B ) Comparative analysis of cell viability in MCF-10A normal cells and cancer cells at their respective cancer cell IC 50 concentrations following 48 h treatment. MCF-10A cells maintained >70% viability across all tested conditions, whereas cancer cells exhibited approximately 50% viability at their IC 50 values, supporting the presence of a potential therapeutic window.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Concentration Assay

Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: Intracellular ROS generation following curcumin (Cur), gemcitabine (Gem), and combination treatment in breast cancer and normal epithelial cells (48 h). Representative DCFH-DA flow cytometry histograms showing intracellular ROS levels in ( A ) MDA-MB-231, ( B ) MCF-7, and ( C ) MCF-10A cells after 48 h treatment with Cur (IC 50 ), Gem (IC 50 ), Cur+Gem, or Cur+Gem with NAC pre-treatment (5 mM, 2 h). DCF fluorescence was detected in the FL1 channel. Cur+Gem treatment induced a rightward shift in fluorescence intensity in both breast cancer cell lines, indicating increased ROS accumulation, whereas NAC pre-treatment reduced this shift. In MCF-10A cells, ROS elevation was limited under identical treatment conditions. Histograms are representative of three independent biological experiments. MFI values are indicated within panels.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Flow Cytometry, Fluorescence

Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: (ΔΨm) alterations following Cur+Gem treatment in breast cancer and normal epithelial cells (48 h). Representative JC-1 flow cytometry dot plots showing mitochondrial membrane potential in MDA-MB-231, MCF-7, and MCF-10A cells after 48 h treatment with Cur+Gem or Cur+Gem with NAC pre-treatment (5 mM, 2 h). JC-1 monomers (FL1) indicate mitochondrial depolarization, whereas JC-1 aggregates (FL2) indicate intact membrane potential. Depo-larized cells were quantified from the lower right quadrant (FL1 + /FL2 − ). Cur+Gem treatment markedly increased the proportion of depolarized cells in MDA-MB-231 (~86%) and MCF-7 (~69%) cells compared to control conditions, where-as NAC pre-treatment reduced depolarization toward control levels. In MCF-10A cells, mitochondrial depolarization following Cur+Gem treatment was limited (~8%) and was reduced after NAC exposure. A minimum of 20,000 events were acquired per sample. Quadrant percentages are indicated within each panel. Data are representative of three inde-pendent biological experiments.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Flow Cytometry, Membrane, Control

Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: Proposed integrative schematic model summarizing the molecular and cellular mechanisms associated with Cur-mediated sensitization to Gem in breast cancer cells. Combined treatment induces intracellular ROS accumulation, which is functionally linked to mitochondrial membrane depolarization (ΔΨm loss) and activation of the intrinsic apoptotic cascade via caspase-9 and caspase-3. The contribution of ROS to this response is supported by NAC-mediated attenuation of mitochondrial depolarization, apoptosis, and synergy scores. Concurrently, transcriptional reprogramming favors a pro-apoptotic shift (↑ BAX, ↑ TP53, ↑ CASP3/9; ↓ BCL2, ↓ NF-κB) and is accompanied by the suppression of angiogenic signaling through VEGFA downregulation. These interconnected redox, mitochondrial, and transcriptional alterations are more pronounced in triple-negative MDA-MB-231 cells, whereas comparatively limited ROS accumulation and mitochondrial disruption are observed in MCF-7 and especially MCF-10A cells under identical conditions, supporting subtype- and cell-selective vulnerability within this in vitro model.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Membrane, Activation Assay, Disruption, In Vitro

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: DOG screening overview. a , Screening overview showing the heatmaps (average of n = 3) of the cell proliferation at 24 h and 72 h for both the MCF10a and MCF7 cells quantified as Ki-67 positive nuclei. The heatmaps corresponding to the cell density and cell area can be found in the supplementary figures. b , Scatter plots showing all data points from one screening (4 DOGs) ranked from low to high. Tissue culture polystyrene (TCP) was used as a control. The 10 % lowest and 10 % highest scoring datapoints are shown on the side and are shown to be significantly different (p < 0.0001). Annotation of black dot with “T” or “L” corresponds to the representative fluorescence images of “Top” and “Low” hits as shown in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Control, Fluorescence

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Top- and low-hit Ki-67-expressing proliferative cells after 72 h of cell culture . Cells were stained for Ki-67 (red), phalloidin (green), and nuclei (blue). Scale bar is 100 μm. White arrows show examples of Ki-67 positive nuclei. The underlying material properties of each image are: MCF10a top hit: S-W 51° 251 MPa, MCF10a low hit: T-S|W 3.8 μm 82° 47 MPa, MCF7 top hit: S-W 63° 42 MPa, MCF7 low hit: T-SW 6 μm 68° 63 MPa.

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Expressing, Cell Culture, Staining

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Scatter plots comparing the effect of each individual physicochemical parameter of the DOG between MCF10a and MCF7 cells. The material parameters (topography, wettability, and stiffness on the same data points, shown with the corresponding color scale) were shown on the biological output (cell proliferation, % Ki-67 positive cells) for the MCF10a vs. the MCF7 cells. Similar plots showing the scatter plots for cell density and cell area can be found in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques:

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: ROIs were chosen that lead to a higher proliferation after 72 h for MCF10a than MCF7 cells. a , Regions of interest (ROIs) are chosen as points furthest away from the dashed line. In orange and blue, points are indicated that show an individual statistical difference (P < 0.05) when comparing the proliferation percentage of MCF7 vs. MCF10a. Orange or blue dots indicate an increase for either MCF7 or MCF10a proliferation, respectively. b , Points from a were grouped and show a significant different in both MCF10a and MCF7 Ki-67 positive cells percentage (p < 0.0001). c , Individual DOG values for each ROI chosen in a are shown. Corresponding fluorescence images for each ROI can be found in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Fluorescence

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: ROIs were chosen from the screening as “positive” when MCF10a Ki-67 % > MCF7 Ki-67 % (labelled as Pos1, Pos2, Pos3, shown in blue) and “negative” when MCF7 Ki-67 % > MCF10a Ki-67 % (labelled as Neg1, Neg2, Neg3, shown in orange). a , Cell experiments were repeated on translational (labelled as Trans) substrates with those specific material properties and compared to the proliferation (% Ki-67 positive cells after 72 h of cell culture) rates from the screening (Screen). b , Cell density after 72 h was compared between the screening and translation findings. c , Fluorescence images of two example ROIs, Pos1 and Neg1. Cells were stained for Ki-67 (red), phalloidin (green), and DAPI (blue). Corresponding fluorescence images for the other ROIs can be found in . The scale bar is 100 μm.

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Cell Culture, Fluorescence, Staining

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Proliferation quantified as % Ki-67 positive cells after 72 h for both MCF7 and MCF10a cells compared between the translational cell experiments on translated ROIs for both low (shown in blue, 1000 cells cm −2 ) and high (shown in orange, 5000 cells cm −2 ) seeding density. a , Individual values for each ROI compared between low and high seeding density. b , All low and high seeding density datapoints from a were grouped and compared for statistical difference (n.s.: p > 0.05, ∗: p < 0.05).

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques:

Journal: American Journal of Cancer Research

Article Title: SIX4 promotes metastasis through STAT3 activation in breast cancer

doi:

Figure Lengend Snippet: SIX4 promotes the activation of the STAT3 signalling pathway in breast cancer. A. MDA-MB-231 cells were silenced with control (Sh-NC) or SIX4 Sh-RNA (no. 1 and 2), and whole-cell extracts were collected for IB analysis of the indicated proteins. B. MCF-7 cells were overexpressed with Vector or SIX4, and whole-cell extracts were collected for IB analysis of the indicated proteins. C, D. IB analysis showed nuclear localization of total STAT3 in those indicated cells. E. HEK293T cells were co-transfected with Flag-SIX4 and HA-STAT3 plasmids, and whole-cell extracts were collected for IP with Flag or HA antibody, followed by IB analysis. F. MDA-MB-231 cells were lysed, then whole-cell extracts were collected for IP with SIX4 or STAT3 antibody, followed by IB analysis. G. Immunofluorescence staining showing co-localization of SIX4 (red) and STAT3 (green) in MDA-MB-231 cells. Scale bar: 10 μm. H. MDA-MB-231 cells were silenced with control (Sh-NC) or SIX4 Sh-RNA#1, whole-cell extracts were collected for IP with STAT3 or lgG antibody, followed by IB analysis of the indicated proteins.

Article Snippet: MDA-MB-231, MDA-MB-468, ZR-75-1, BT-549, MCF-7, MCF-10A and HEK293T cells were obtained from Shanghai GENECHEM, Co., Ltd.

Techniques: Activation Assay, Control, Plasmid Preparation, Transfection, Immunofluorescence, Staining