Journal: Cancers

Article Title: High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer

doi: 10.3390/cancers14010002

Figure Lengend Snippet: Target control. MCF-7 ( A ) and SkBr3 ( B ) cells were treated with 10 µM P8-D6 (fluorophore: 462Ex/530Em) or control (PBS) for 10 h. P8-D6 were localized in vitro. ( i ): Cells were stained by CellTracker TM Deep Red Dye and Hoechst 33342. ( ii ) After fixation, topoisomerase I were detected. Fluorescence images show the fluorophore P8-D6 in green, membrane staining ( i ) or topoisomerase expression ( ii ) in red and nucleus staining in white. Scale bars, 50 µm. ( C ) Fluorescence intensity of P8-D6 in the nuclei was compared to PBS control. ( D ) BC cell lines and primary cells were lysed and protein expression was analyzed using western blot. HSP 90 was used as loading control.

Article Snippet: MCF-7 (1000/well), SkBr3 (1000/well), MDA-MB231 (2500/well), MDA-MB468 (500/well), BT-20 (18,750/well) and UF-182 (20,000/well) cells were seeded onto a 96-well Ultra-Low Attachment plate (Corning #4520).

Techniques: Control, In Vitro, Staining, Fluorescence, Membrane, Expressing, Western Blot

Journal: Cancers

Article Title: High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer

doi: 10.3390/cancers14010002

Figure Lengend Snippet: Antitumor responses in BC 2D monolayers. MCF-7, SkBr3, MDA-MB231, MDA-MB468, BT-20 (cell line) and UF-182 (primary cells) were treated with P8-D6, cisplatin, etoposide, topotecan and PBS as control. The relative caspase activity representing the rate of apoptosis were measured 48 h after treatment in MCF-7 ( A ), SkBr3 ( B ), MDA-MB231 ( C ), MDA-MB468 ( D ), BT-20 ( E ) and UF-182 ( F ) cells. Additionally, the anti-proliferative effects in the cells were visualized by microscopy after 24 h treatment. Scale bars, 50 µm ( G ). Heat map presents the IC50 values calculated by viability ( H ). P8-D6 was compared to cisplatin, epirubicin and topotecan. Data are means + SD one-way ANOVA, * ( p < 0.05), ** ( p < 0.01), *** ( p < 0.001), **** ( p < 0.0001), ns (non-significant).

Article Snippet: MCF-7 (1000/well), SkBr3 (1000/well), MDA-MB231 (2500/well), MDA-MB468 (500/well), BT-20 (18,750/well) and UF-182 (20,000/well) cells were seeded onto a 96-well Ultra-Low Attachment plate (Corning #4520).

Techniques: Control, Activity Assay, Microscopy

Journal: Cancers

Article Title: High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer

doi: 10.3390/cancers14010002

Figure Lengend Snippet: Growth changes in BC spheroids. For 3D culture, MCF-7, SkBr3, MDA-MB231, MDA-MB468, BT-20 (cell line) and UF-182 (primary cells) cells were cultured in ULA plates for 96 h. Subsequently, spheroids were treated with P8-D6 (10 µM, 1 µM, 0.5 µM, 0.1 µM), topotecan (10 µM, 1 µM) and PBS for 48 h. For the monitoring of growth changes and morphological changes, spheroids were imaged every 24 h by microscopy. MCF-7 ( A ), SkBr3 ( B ), MDA-MB231 ( C ), MDA-MB468 ( D ), BT-20 ( E ) and UF-182 ( F ). Scale bars, 500 µm.

Article Snippet: MCF-7 (1000/well), SkBr3 (1000/well), MDA-MB231 (2500/well), MDA-MB468 (500/well), BT-20 (18,750/well) and UF-182 (20,000/well) cells were seeded onto a 96-well Ultra-Low Attachment plate (Corning #4520).

Techniques: Cell Culture, Microscopy

Journal: Cancers

Article Title: High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer

doi: 10.3390/cancers14010002

Figure Lengend Snippet: Apoptosis induction in BC spheroids. For 3D culture, MCF-7, SkBr3, MDA-MB231, MDA-MB468, BT-20 (cell line) and UF-182 (primary cells) cells were cultured in ULA plates for 96 h. Subsequently, spheroids were treated with P8-D6 (10 µM, 1 µM, 0.5 µM, 0.1 µM), topotecan (10 µM, 1 µM) and PBS for 48 h. After treatment, the viability and caspase activity were analyzed in MCF-7 ( A ), SkBr3 ( B ), MDA-MB231 ( C ), MDA-MB468 ( D ), BT-20 ( E ) and UF-182 ( F ) spheroids. Data are means + SD ( n = 3) one-way ANOVA, * ( p < 0.05), ** ( p < 0.01), *** ( p < 0.001), **** ( p < 0.0001), ns (non-significant).

Article Snippet: MCF-7 (1000/well), SkBr3 (1000/well), MDA-MB231 (2500/well), MDA-MB468 (500/well), BT-20 (18,750/well) and UF-182 (20,000/well) cells were seeded onto a 96-well Ultra-Low Attachment plate (Corning #4520).

Techniques: Cell Culture, Activity Assay

Journal: Cancers

Article Title: High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer

doi: 10.3390/cancers14010002

Figure Lengend Snippet: Cell toxicity, live-dead staining and morphological changes in BC spheroids. For 3D culture, MCF-7, SkBr3, MDA-MB231, MDA-MB468, BT-20 (cell line) and UF-182 (primary cells) cells were cultured in ULA plates for 96 h. Subsequently, spheroids were treated with P8-D6 (10 µM, 1 µM, 0.5 µM, 0.1 µM), topotecan (10 µM, 1 µM) and PBS for 48 h. After treatment, the spheroids were stained with propidium iodide (red), calcein-AM (green) and Hoechst 33342 (blue) and imaged using NYONE ®® Scientific. MCF-7 ( A ), SkBr3 ( B ) and UF-182 ( C ). Scale bars, 500 µm. ( D ) During treatment, the cell toxicity was measured by fluorescence microscopy using CellTox™ Green (24 h, 48 h). Scale bars, 500 µm. These fluorescence signals were quantified (fluorescence intensity RFU) and presented in a heat map ( E ). ( F ) The P8-D6- (1 μM) or PBS-treated SkBr3 and UF-182 spheroids (48 h) were analyzed by scanning electron microscopy (SEM). Scale bars, 20 µm.

Article Snippet: MCF-7 (1000/well), SkBr3 (1000/well), MDA-MB231 (2500/well), MDA-MB468 (500/well), BT-20 (18,750/well) and UF-182 (20,000/well) cells were seeded onto a 96-well Ultra-Low Attachment plate (Corning #4520).

Techniques: Staining, Cell Culture, Fluorescence, Microscopy, CellTox Assay, Electron Microscopy

Journal: Cell reports

Article Title: Breast tumor stiffness instructs bone metastasis via maintenance of mechanical conditioning

doi: 10.1016/j.celrep.2021.109293

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: After mechanical preconditioning, SUM159, SKBR3 and MS-SKBR3.1 cells expressing iRFP-LifeAct were trypsizined from their hydrogels and plated onto No. 1.5 glass MatTek dishes which had been pre-treated overnight with DMEM + 10% FBS, and then incubated for 10 hours to ensure maximal spreading before analysis (verified by size equilibrium).

Techniques: Recombinant, Membrane, Enzyme-linked Immunosorbent Assay, Software

Journal: Theranostics

Article Title: Investigating the Intracellular Behaviors of Liposomal Nanohybrids via SERS: Insights into the Influence of Metal Nanoparticles

doi: 10.7150/thno.21173

Figure Lengend Snippet: Bright field image (A), SERS mapping (B) and merged image (C) of SKBR3 cells incubated with nanohybrids for 5 h. (D) SERS spectrum acquired from the SKBR3 cells. Scale bars for confocal images are 10 μm.

Article Snippet: After nanohybrids-loaded SKBR3 cells were supplemented with fresh culture medium, exocytosis prevailed gradually, leading to a decrease in the content of nanohybrids within SKBR3 cells as well as a reduction in the intensity of SERS signal.

Techniques: Incubation

Journal: Theranostics

Article Title: Investigating the Intracellular Behaviors of Liposomal Nanohybrids via SERS: Insights into the Influence of Metal Nanoparticles

doi: 10.7150/thno.21173

Figure Lengend Snippet: Colocalization between nanoparticles (labeled by SERS, red) and lipid bilayers (labeled by fluorescence, green) for 4 h (A-C), 8 h (D-F) and 12 h (G-I); 580-610 nm light was collected for fluorescence imaging of DiI and 800-1600 cm -1 Raman scattered light was collected for SERS imaging. (J) Average SERS spectrum of a single SKBR3 cell incubated with the nanohybrids for different times. (K) Colocalization quantification (>0.6 indicates substantial colocalization). Scale bars for confocal images are 10 μm.

Article Snippet: After nanohybrids-loaded SKBR3 cells were supplemented with fresh culture medium, exocytosis prevailed gradually, leading to a decrease in the content of nanohybrids within SKBR3 cells as well as a reduction in the intensity of SERS signal.

Techniques: Labeling, Fluorescence, Imaging, Incubation

Journal: Theranostics

Article Title: Investigating the Intracellular Behaviors of Liposomal Nanohybrids via SERS: Insights into the Influence of Metal Nanoparticles

doi: 10.7150/thno.21173

Figure Lengend Snippet: Effect of inhibitors on uptake of SKBR3 cells incubated with nanohybrids probed by SERS. In the intracellular experiments, 580-610 nm light was collected for fluorescence imaging of DiI (green) and 800-1600 cm -1 Raman scattered light was collected for SERS imaging (red). Scale bars for confocal images are 10 μm.

Article Snippet: After nanohybrids-loaded SKBR3 cells were supplemented with fresh culture medium, exocytosis prevailed gradually, leading to a decrease in the content of nanohybrids within SKBR3 cells as well as a reduction in the intensity of SERS signal.

Techniques: Incubation, Fluorescence, Imaging

Journal: Theranostics

Article Title: Investigating the Intracellular Behaviors of Liposomal Nanohybrids via SERS: Insights into the Influence of Metal Nanoparticles

doi: 10.7150/thno.21173

Figure Lengend Snippet: Confocal microscopy of SERS labeled nanohybrids (red) and fluorescence staining of organelle markers (green). Makers are DiI (membranes, A-C) and lyso-tracker green (lysosomes, D-F): 500-540 nm light was collected for the fluorescence imaging of Lysotracker Green, 580-610 nm light was collected for fluorescence imaging of DiI and 800-1600 cm -1 Raman scattered light was collected for SERS imaging. (G) Average SERS intensity of a single SKBR3 cell incubated with the nanohybrids for 4, 8 and 12 h. (H) Colocalization coefficients between nanohybrids and membranes (or lysosomes). Scale bars for confocal images are 10 μm.

Article Snippet: After nanohybrids-loaded SKBR3 cells were supplemented with fresh culture medium, exocytosis prevailed gradually, leading to a decrease in the content of nanohybrids within SKBR3 cells as well as a reduction in the intensity of SERS signal.

Techniques: Confocal Microscopy, Labeling, Fluorescence, Staining, Imaging, Incubation

Journal: Theranostics

Article Title: Investigating the Intracellular Behaviors of Liposomal Nanohybrids via SERS: Insights into the Influence of Metal Nanoparticles

doi: 10.7150/thno.21173

Figure Lengend Snippet: Confocal microscopy of NBD labeled liposomes (green) and fluorescence staining of organelle markers (red). Makers are DiI (membranes, A-C) and lysotracker red (lysosomes, D-F): 520-550 nm light was collected for fluorescence imaging of NBD, 580-610 nm light was collected for fluorescence imaging of DiI and 570-620 nm light was collected for fluorescence imaging of Lysotracker Red. (G) Average fluorescence intensity of a single SKBR3 cell incubated with the nanohybrids for 4, 8 and 12 h. (H) Colocalization coefficients between liposomes and membranes (or lysosomes). Scale bars for confocal images are 10 μm.

Article Snippet: After nanohybrids-loaded SKBR3 cells were supplemented with fresh culture medium, exocytosis prevailed gradually, leading to a decrease in the content of nanohybrids within SKBR3 cells as well as a reduction in the intensity of SERS signal.

Techniques: Confocal Microscopy, Labeling, Liposomes, Fluorescence, Staining, Imaging, Incubation