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: Journal of Breast Cancer

Article Title: Enhancing the Effects of Low Dose Doxorubicin Treatment by the Radiation in T47D and SKBR3 Breast Cancer Cells

doi: 10.4048/jbc.2013.16.2.164

Figure Lengend Snippet: Effect of irrdiation alone and in combination with doxorubicin on viability of T47D (A) and SKBR3 (B) cells. The cells were treated with 1 µM doxorubicin and irradiated with doses of 100, 150, and 200 cGy. IR=irradiation; DOX=doxorubicin.

Article Snippet: The human breast cancer cell lines, SKBR3 and T47D (purchased from National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran), were grown in RPMI-1640 media supplemented with 10% heat-inactivated (50°C, 30 minutes) fetal bovine serum, penicillin (100 U/mL), streptomycin (100 μg/mL), and amphotericin B (0.25 μg/mL) at 5% CO 2 and 95% air in a humidified 37°C incubator.

Techniques: Irradiation

Journal: Journal of Breast Cancer

Article Title: Enhancing the Effects of Low Dose Doxorubicin Treatment by the Radiation in T47D and SKBR3 Breast Cancer Cells

doi: 10.4048/jbc.2013.16.2.164

Figure Lengend Snippet: Viability of T47D and SKBR3 cell lines following combined treatment with 1 µM doxorubicin and 100,150, 200 cGy irradiation. IR=irradiation; DOX=doxorubicin.

Article Snippet: The human breast cancer cell lines, SKBR3 and T47D (purchased from National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran), were grown in RPMI-1640 media supplemented with 10% heat-inactivated (50°C, 30 minutes) fetal bovine serum, penicillin (100 U/mL), streptomycin (100 μg/mL), and amphotericin B (0.25 μg/mL) at 5% CO 2 and 95% air in a humidified 37°C incubator.

Techniques: Irradiation

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: Molecular Cancer

Article Title: Exosome-mediated lncRNA AFAP1-AS1 promotes trastuzumab resistance through binding with AUF1 and activating ERBB2 translation

doi: 10.1186/s12943-020-1145-5

Figure Lengend Snippet: Knockdown of AFAP1-AS1 reversed trastuzumab resistance in vivo. a Representative images of the xenograft after intraperitoneal injection with once every 2 days 3 mg/kg trastuzumab treatment for 3 weeks in sh-NC or sh-AFAP1-AS1 cell groups. b Tumor sizes were calculated after injection every 2 days, *** P < 0.001 compared to sh-NC group. c Immunohistochemistry analysis revealed that the tumors developed from sh-AFAP1-AS1 cells displayed lower HER-2 staining than the sh-NC group. d-e Luciferase flux count ( d ) along with the visible number of metastases in lung ( e ) were significantly less in sh-AFAP1-AS1 group compared to sh-NC group. f Visualization of the HE-stained lung tissues from different sections

Article Snippet: To experimentally construct lung metastases, SKBR-3-TR cells that had been transfected to stably express firefly luciferase (Xenogen Corporation, CA, USA) were infected with lentiviruses carrying control shRNA or sh-AFAP1-AS1.

Techniques: Knockdown, In Vivo, Injection, Immunohistochemistry, Staining, Luciferase