Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: ROS generation detection in vitro . (A) The release of singlet oxygen from each drug was detected in vitro under dark conditions. Data were presented as the mean ± SD ( n = 3). (B) Detection of singlet oxygen release of each drug under 660 nm laser irradiation at a power density of 280 mW cm -2 was performed in vitro . Data were presented as the mean ± SD ( n = 3). (C) ROS generation was observed in 4T1 cells treated with each medication while being exposed to laser irradiation at a wavelength of 660 nm and intensity of 280 mW cm −2 . Bright: Bright field. DCF: Green fluorescence represents intracellular ROS. Merge: Superimpose the image. Scale Bar = 100 μm.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vitro, Irradiation, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Evaluation of mitochondrial targeting function. (A) Confocal images of the mitochondrial sites of TPPOH 2 , CTT 2 , CTT 2 P, and CTT 2 P@B NPs in 4T1 cells. Mito-Tracker Green was used to stain the mitochondria in the green channel. The red channel was derived from the emission of the photosensitizer fraction (PS) itself. Merge stands for superimposed image. The Green and red curves in the Plot Profile represent the gray value of Mito-Tracker Green and PS, respectively. Scale bar = 20 μm. (B) Flow cytometry JC-1 method was used to analyze the mitochondrial function of cells treated with different drugs. Red fluorescence: normal mitochondria (J-aggregate); Green fluorescence: depolarized mitochondria (J-monomer).

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: Staining, Derivative Assay, Flow Cytometry, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Evaluation of cell death in vitro through cytotoxicity assessment and examination of apoptosis and necrosis. (A) The in vitro cytotoxicity of 4T1 cells treated with CPT, TPPOH 2 , CTT 2 , CTT 2 P, and CTT 2 P@B NPs in the dark was assessed using the CCK-8 assay. Data were presented as the mean ± SD ( n = 5). ** p < 0.01, **** p < 0.0001. (B) The in vitro cytotoxicity of 4T1 cells treated with TPPOH 2 , CTT 2 , CTT 2 P and CTT 2 P@B NPs under laser irradiation (660 nm, 280 mW cm −2 ) was determined using the CCK-8 assay. Data were presented as the mean ± SD ( n = 5). ** p < 0.01, **** p < 0.0001. (C) Cell apoptosis and necrosis were analyzed using flow cytometry with Annexin V-FITC/PI double staining following treatment with various drugs at a concentration of 5 μM.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vitro, CCK-8 Assay, Irradiation, Flow Cytometry, Double Staining, Concentration Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Biodistribution in vivo . (A) Blood compatibility test of CPT, TPPOH 2 , CTT 2 , CTT 2 P, CTT 2 P@B NPs. Data were presented as the mean ± SD ( n = 3). *** p < 0.001, **** p < 0.0001. (B) Time-lapse live fluorescence imaging of mice with 4T1 tumors following the administration of free CTT 2 P and CTT 2 P@B NPs via intravenous injection. (C) Fluorescent images of major organs and tumors were obtained 24 h after injection, using ex vivo methods. (D) The mean fluorescence intensity of each organ and tumor was used to determine the biodistribution of free CTT 2 P and CTT 2 P@B NPs in mice. Data were presented as the mean ± SD ( n = 3). * p < 0.05.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vivo, Fluorescence, Imaging, Injection, Ex Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: In vivo anti-tumor study of each drug in 4T1 tumor-bearing mice. (A) Tumor images of different drug administration treatments after the antitumor study. (B) During the administration, the growth of tumors in mice was observed in each group receiving treatment. Data were presented as the mean ± SD ( n = 5), ** p < 0.01. (C) The weight of mice in each treatment group was monitored throughout the administration period. Data were presented as the mean ± SD ( n = 5).

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Investigation of the effects of each medication on mice with 4T1 tumors through pathological examination. (A) After administering various medications, the major organs (including the heart, liver, spleen, lung, and kidney) were subjected to H&E staining. Scale bar = 50 μm. (B) After administering various medications, tumors were subjected to H&E staining and TUNEL staining. Scale bar = 50 μm.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: Medications, Staining, TUNEL Assay

Journal: Molecular Pharmaceutics

Article Title: Non-Invasive Detection of Passively Targeted Poly(ethylene glycol) Nanocarriers in Tumors

doi: 10.1021/mp2003913

Figure Lengend Snippet: Passive tumor distribution of PEG nanocarriers measured non-invasively using a SkinSkan® spectrofluorometer. Nanocarriers (0.5 mM) were intravenously administered to 4T1 tumor-bearing balb/c mice and fluorescence spectra were collected from tumor and a contralateral skin (λexc.: 480 nm; λem.: 515 nm for 10–30 kDa and 520 nm for 40–60 kDa). Each column and error bar represents the mean±SD (n = 5–7). Individual comparisons between the groups were determined by Student’s t-test using the Microsoft Excel 2008. *: p<0.05, tumor vs. corresponding dermal control site.

Article Snippet: The 4T1 murine breast cancer cell line was a generous gift from Dr. Michael Reiss, The Cancer Institute of New Jersey (New Brunswick, NJ).

Techniques: Fluorescence, Control

Journal: Molecular Pharmaceutics

Article Title: Non-Invasive Detection of Passively Targeted Poly(ethylene glycol) Nanocarriers in Tumors

doi: 10.1021/mp2003913

Figure Lengend Snippet: (A) Non-invasive images showing the passive tumor distribution of PEG nanocarriers. The images were obtained on an IVIS® 100 imaging system using the excitation and emission filters corresponding to green fluorescent protein (GFP). The first two animals (balb/c mice with 4T1 tumor), starting from the left, were untreated (controls), whereas the remaining three animals were administered nanocarriers (0.5 mM) intravenously; (B) Regions of interest for tumor and control areas are shown in a mouse injected with 40 kDa nanocarrier for average efficient quantitation; and (C) Plots of average fluorescence from tumor and control site at skin’s surface against time. Each point represents the mean±SD (n = 3). Individual comparisons between groups were determined by Student’s t-test using the Microsoft Excel 2008. *: p<0.01, tumor vs. corresponding control site.

Article Snippet: The 4T1 murine breast cancer cell line was a generous gift from Dr. Michael Reiss, The Cancer Institute of New Jersey (New Brunswick, NJ).

Techniques: Imaging, Control, Injection, Quantitation Assay, Fluorescence

Journal: Molecular Pharmaceutics

Article Title: Non-Invasive Detection of Passively Targeted Poly(ethylene glycol) Nanocarriers in Tumors

doi: 10.1021/mp2003913

Figure Lengend Snippet: Ex vivo distribution studies: (A) Tissue distribution at 24 hrs; (B) Tumor distribution at 24 and 96 hrs; and (C) Plasma distribution at 24 and 96 hrs. PEG nanocarriers (0.5 mM) were intravenously administered to balb/c mice bearing 4T1 tumors and animals were euthanized to collect tissues. Each column and error bar represents the mean±SD (n = 5–7). The statistical analyses were carried out using GraphPad Prism v.4 as follows: (A) Two-way ANOVA and individual comparison between the groups were determined using Bonferroni posttests. The statistically significant groups (p<0.05, 60 kDa vs. other nanocarriers) are marked as * (10 kDa); & (20 kDa); # (30 kDa); and + (40 kDa); (B) One-way ANOVA and individual comparison between the groups were determined using Tukey posttests. The statistically significant groups (p<0.05, 60 kDa vs. other nanocarriers are marked as * (10, 20 or 30 kDa); and # (40 kDa at 96 hrs); (C) One-way ANOVA and comparison between the groups were determined using Tukey posttests. The statistically significant groups (p<0.05, 60 kDa vs. other nanocarriers) are denoted by * (10 or 20 kDa); & (30 kDa); # (40 kDa); and + (nanocarriers at 96 hrs).

Article Snippet: The 4T1 murine breast cancer cell line was a generous gift from Dr. Michael Reiss, The Cancer Institute of New Jersey (New Brunswick, NJ).

Techniques: Ex Vivo, Clinical Proteomics, Comparison

Journal: Oncogenesis

Article Title: The phosphorylated prodrug FTY720 is a histone deacetylase inhibitor that reactivates ERα expression and enhances hormonal therapy for breast cancer

doi: 10.1038/oncsis.2015.16

Figure Lengend Snippet: FTY720-P is produced in the nucleus of breast cancer cells by SphK2. Breast cancer cell lines, murine 4T1 ( a, b ), human MDA-MB-231 ( c, d ) and human MCF7 ( f ) were treated with 5 μ M FTY720. ( a, c ) Nuclear levels of FTY720-P and S1P were determined by liquid chromatography, electrospray ionization/tandem mass spectrometry (LC-ESI-MS/MS) at the indicated times. Equal amounts of protein from nuclear and cytosolic fractions were analyzed by immunoblotting with SphK2 antibody. Antibodies against histone H3 or laminA/C and tubulin were used as nuclear and cytosol markers. ( b , d ) Total intracellular and secreted FTY720-P were determined in 4T1 cells after 8 h and MDA-MB-231 cells after 6 h of FTY720 treatment, respectively. MDA-MB-231 cells ( e ) and MCF7 cells ( g ) transfected with vector, SphK2 or catalytically inactive SphK2 G212E (ciSphK2) were treated with vehicle or 5 μ M FTY720 for 6 and 24 h, respectively. Nuclear levels of FTY720-P and S1P were determined by LC-ESI-MS/MS. Data are mean±s.d. * P <0.005 compared with vector; # P <0.005 compared with vehicle. Equal expression of nuclear SphK2 was confirmed by immunoblotting.

Article Snippet: Human breast cancer cells MCF7 and MDA-MB-231 (ATCC, Manassas, VA, USA) and murine 4T1 breast cancer cells (Caliper Life Sciences, Waltham, MA, USA) were cultured and transfected with vector, SphK2 or catalytically inactive SphK2 G212E as previously described.

Techniques: Produced, Liquid Chromatography, Mass Spectrometry, Tandem Mass Spectroscopy, Western Blot, Transfection, Plasmid Preparation, Expressing

Journal: Oncogenesis

Article Title: The phosphorylated prodrug FTY720 is a histone deacetylase inhibitor that reactivates ERα expression and enhances hormonal therapy for breast cancer

doi: 10.1038/oncsis.2015.16

Figure Lengend Snippet: Nuclear FTY720-P enhances specific histone acetylations in breast cancer cells. MCF7 cells ( a ) and 4T1 cells ( b ) were treated with FTY720 (5 μ M ) for the indicated times. Histone acetylations in nuclear extracts were detected by immunoblotting with antibodies to specific histone acetylation sites. ( c ) Purified nuclei from naive MCF7 cells were incubated for the indicated times with vehicle, S1P (1 μ M ) or FTY720-P (1 μ M ) and histone acetylations determined. ( d , e ) Purified nuclei were isolated from MCF7 cells transfected with vector, SphK2 or ciSphK2 and treated with FTY720 (1 μ M ) for 15 min. ( f , g ) Purified nuclei were isolated from MCF7 cells transfected with siControl or siSphK2 and incubated with the indicated concentrations of FTY720 for 15 min. Histone acetylations were determined by immunoblotting ( d , f ) and levels of FTY720-P by liquid chromatography, electrospray ionization/tandem mass spectrometry (LC-ESI-MS/MS) ( e , g ). * P <0.05. ( h, i ) Naive MCF7 cells were treated with vehicle, FTY720-P (100 n M ), FTY720 (1 μ M ) or SAHA (2 μ M ) for 2 h, nuclear extracts were analyzed by western blotting with the indicated antibodies ( h ) and HDAC activity measured and expressed as arbitrary fluorescence units (AFU) ( i ). * P <0.001.

Article Snippet: Human breast cancer cells MCF7 and MDA-MB-231 (ATCC, Manassas, VA, USA) and murine 4T1 breast cancer cells (Caliper Life Sciences, Waltham, MA, USA) were cultured and transfected with vector, SphK2 or catalytically inactive SphK2 G212E as previously described.

Techniques: Western Blot, Purification, Incubation, Isolation, Transfection, Plasmid Preparation, Liquid Chromatography, Mass Spectrometry, Tandem Mass Spectroscopy, Activity Assay, Fluorescence

Journal: Oncogenesis

Article Title: The phosphorylated prodrug FTY720 is a histone deacetylase inhibitor that reactivates ERα expression and enhances hormonal therapy for breast cancer

doi: 10.1038/oncsis.2015.16

Figure Lengend Snippet: FTY720 induces ERα expression in ERα-negative human and murine breast cancer cells and sensitizes them to tamoxifen. 4T1 ( a ) and MDA-MB-231 ( b ) cells were treated with FTY720 (5 μ M ) or SAHA (1 μ M ) for 24 h. ERα, PR and ERβ mRNA levels were determined by quantitative real-time PCR (QPCR) and normalized to GAPDH. ( c ) Proteins in MDA-MB-231 nuclear extracts were analyzed by immunoblotting with the indicated antibodies. LaminA/C was used as a loading control. ( d ) MDA-MB-231 cells were subjected to chromatin immunoprecipitation (ChIP) analyses with antibodies to H3-ac, H3 or normal rabbit IgG, as indicated. The precipitated DNA was analyzed by real-time PCR with primers amplifying the core promoter sequence of the ERα gene. Relative binding to the promoter is expressed as fold enrichment compared with input. Data are mean±s.d. * P <0.003 compared with vehicle. ( e ) MDA-MB-231 cells were treated with FTY720 (1 n M ) without or with 10 n M E2 for the indicated days and cell proliferation was determined by WST assay. ( f , g ) MDA-MB-231 cells ( f ) or 4T1 cells ( g ) were treated with the indicated concentrations of TAM or FTY720, or with 2.5 μ M FTY720 with increasing concentrations of TAM for 48 h and cell proliferation determined. Data are expressed as % of untreated control.

Article Snippet: Human breast cancer cells MCF7 and MDA-MB-231 (ATCC, Manassas, VA, USA) and murine 4T1 breast cancer cells (Caliper Life Sciences, Waltham, MA, USA) were cultured and transfected with vector, SphK2 or catalytically inactive SphK2 G212E as previously described.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Chromatin Immunoprecipitation, Sequencing, Binding Assay, WST Assay

Journal: Oncogenesis

Article Title: The phosphorylated prodrug FTY720 is a histone deacetylase inhibitor that reactivates ERα expression and enhances hormonal therapy for breast cancer

doi: 10.1038/oncsis.2015.16

Figure Lengend Snippet: FTY720 reduces breast tumor growth and enhances anticancer effectiveness of TAM in ERα-negative 4T1 syngeneic xenografts. 4T1 cells were surgically implanted into the second mammary fat pads under direct vision. Tumor-bearing mice were randomized into five groups 2 days after implantation and then treated with vehicle, FTY720 (1 mg/kg), TAM (25 mg/kg), FTY720 plus TAM or SAHA (intraperitoneal (i.p.) 20 mg/kg) plus TAM by gavage daily till day 15 ( n =8). ( a ) Tumor volumes were measured daily. (Insert) Tumor volumes on day 15. ( b ) Representative tumors. * P <0.01, # P <0.05 compared with vehicle. ( c , d ) Immunohistochemical staining of tumor sections for TUNEL ( c ) and ERα ( d ). Scale bar: 20 μm. Quantifications of TUNEL-positive cells and ERα intensity are shown. * P <0.05 compared with vehicle or TAM. ( e ) HDAC activity in nuclear extracts of tumors was determined and expressed as arbitrary fluorescence units. ( f ) Expression of ERα in the tumors was analyzed by quantitative real-time PCR (QPCR) and normalized to Gapdh . ( g ) Nuclear extract proteins were analyzed by western blotting with the indicated antibodies. Histone H3 was used as loading control. Data are mean±s.e.m. * P <0.01 compared with vehicle or TAM.

Article Snippet: Human breast cancer cells MCF7 and MDA-MB-231 (ATCC, Manassas, VA, USA) and murine 4T1 breast cancer cells (Caliper Life Sciences, Waltham, MA, USA) were cultured and transfected with vector, SphK2 or catalytically inactive SphK2 G212E as previously described.

Techniques: Immunohistochemical staining, Staining, TUNEL Assay, Activity Assay, Fluorescence, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control