Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Growth rates of the murine adenocarcinoma 4T1 cell line and its sublclones expressing luciferase 4T1luc2D6 and 4T1luc2. The time course of the proliferation of the parental 4T1, and 4T1luc2D6 and 4T1luc2 (PerkinElmer) clones as determined using Nikon Biostation CT (Nikon, Japan) with STDV ( A ); Bioluminescence from 4T1luc2 and 4T1luc2D6 cells in culture assessed by bioluminescent imaging (Spectrum CT, Perkin Elmer) ( B ); Average level of bioluminescence of 4T1luc2 and 4T1luc2D6 cells (photons/cell/sec) ( C ). Results from five independent measurements. No significant difference between any of the analysed parameters (p > 0,05; Mann Whitney test).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Expressing, Luciferase, Clone Assay, Imaging, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Growth in BALB/c mice (n = 5–6 per group) of primary tumors induced by the implantation of 4T1, 4T1luc2 and 4T1lucD6 cells (see Methods for description). Growth curves obtained by MRI visualize average tumor volume in cubic mm, with STDV ( A ); Growth of primary tumors induced by implantation of 4T1luc2 and 4T1lucD6 cells monitored by bioluminescent imaging; curves depict an average photon flux from the tumor per sec, with STDV ( B ); Statistical comparison of the tumor sizes evaluated by MRI ( C ). Median size of 4T1lucD6 tumors is significantly lower (p = 0,017), and of 4T1luc2 tumors tend to be lower than of 4T1 tumors, although the difference is not significant (p = 0,17). Day of implantation is counted as day 0. Statistical comparisons are done using Kruskal Wallis and Mann-Whitney tests (Statistica AXA 10.0).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Imaging, Comparison, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Comparison of the luciferase activity measured as the intensity of luminescence (in arbitrary units a.u.; Enspire, Perkin Elmer) in the original 4T1luc2 and 4T1luc2D6 cell lines and in cell cultures prepared from the 4T1luc2 and 4T1luc2D6 tumors by the experimental end-point. Luciferase activity in the 4T1luc2 and 4T1luc2D6 cells prior to implantation ( A ); Luciferase activity in primary cell cultures prepared from tumors formed in BALB/c mice by implantation of 4T1luc2D6 (n = 3; 6 tumors) ( B ) or 4T1luc2 cells (n = 3; 6 tumors) ( C ); Recalculation of the average luciferase activity per cell with STDV ( D ). Individual tumors are coded by the cell line, mouse and tumor numbers, for example “4T1luc2D6 n10 r1” designate a tumor caused by implantation of cell line 4T1luc2D6 into mouse No. 10 and refer to sample No. 1 from one of the tumors. Curves in panels B and D show dependence of luciferase activity on the number of cells used in the assay; data represent the average of three parallel measurements. Statistical comparisons are done using Mann-Whitney test (Statistica AXA 10.0); **p < 0,01, and *p < 0,05.

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Comparison, Luciferase, Activity Assay, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: In vitro IFN-gamma response of lymphocytes of BALB/c mice implanted with Luc-expressing 4T1luc2D6 or 4Tl1luc2, or parental 4T1 cells, to stimulation with peptide GFQSMYTFV representing an immunodominant CTL epitope of luciferase (LucP) assessed using IFN-γ ELISpot (Mabtech). IFN-γ response to LucP by the experimental end-point assessed as the average number of IFN-γ spot forming cells per mln splenocytes (SFC/mln) with STDV; *p < 0.05, IFN-γ response to LucP in BALB/c mice implanted with 4T1luc2D6 and 4T1luc2 cells compared to IFN-γ response exhibited by naïve or 4T1-implanted mice (Mann-Whitney test) ( A ); Development of cellular immune response to LucP in mice implanted with 5000 4T1luc2 cells, on days 6, 9 and 23 post implantation; *p < 0.05, IFN-γ response to LucP in BALB/c mice implanted with 4T1luc2 cells on days 9 and 23 post implantation compared to IFN-γ response exhibited by naïve mice or mice implanted by 4T1luc2 and assessed on day 6 post implantation (Friedman ANOVA and Kendall Coeff. of Concordance) ( B ). Red line indicates the cut-off for the specific IFN-γ response, defined as an average number of lymphocytes producing IFN-γ in response to stimulation with LucP ± 3 STDV per mln lymphocytes. Cut-off was established in independent in vitro tests done on splenocytes of naïve mice (n = 5).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: In Vitro, Expressing, Luciferase, Enzyme-linked Immunospot, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Intradermal Luc DNA immunization followed by electroporation (EP) with driving pulses of 100 V makes BALB/c mice resistant to initiation of 4T1luc2 tumors. BALB/c mice were immunized by plasmid encoding firefly luciferase pVaxLuc delivered by intradermal injections followed by EP using multineedle (Mn) electrodes mounted on Dermavax (Cellectis) (n = 4) or CUY21EditII (BEX Ltd) electroporators at 100 V (n = 6) or 50 V (n = 4). Immunization (red circles) and 4T1luc2 implantation sites (blue circles) visualized by in vivo imaging (Spectrum CT) on days 1 and 6 post tumor cell implantation (43 and 48 days of the immunization cycle). Mice immunized with Luc DNA (A, upper panel) versus empty vector (A, lower panel). Text boxes demonstrate total flux from the respective regions of interest (ROI) (photons/sec) ( A ); Quantification of bioluminescence emission (BLI; photon flux/sec) from immunization sites illustrates the efficacy of Luc gene transfer and expression. From day 5, Luc DNA immunized mice electroporated at 100 V emitted stronger bioluminescence than mice electroporated at 50 V (p < 0,05; Mann Whitney test), no bioluminescence from immunization sites in control mice receiving pVax1 ( B ); Quantification of bioluminescence (photon flux/sec) from 4T1luc2 implantation sites in mice immunized with Luc DNA and electroporated with DermaVax/Mn at 100 V (n = 4), BEX/Mn at 100 V (n = 6), BEX/Mn at 50 V (n = 4), and vector immunized mice (n = 3) on days 0 to 6 post challenge ( C ); Difference (in %) in BLI from 4T1luc2 implantation sites after six days of tumor growth in mice immunized with Luc DNA with EP at 100 V (DermaVax/Mn + BEX/Mn groups, 10 animals, dubbed “Luc, 100 V”), or 50 V (BEX/Mn, 4 animals, dubbed “Luc, 50 V”), vector immunized (n = 3) and naive mice (n = 5, from an independent experiment) ( D ). *p < 0,05; **p < 0,1 (Statistica AXA 10.0).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Electroporation, Plasmid Preparation, Luciferase, In Vivo Imaging, Expressing, MANN-WHITNEY, Control

Journal: Scientific Reports

Article Title: Luciferase Expression Allows Bioluminescence Imaging But Imposes Limitations on the Orthotopic Mouse (4T1) Model of Breast Cancer

doi: 10.1038/s41598-017-07851-z

Figure Lengend Snippet: Cellular immune response to peptide GFQSMYTFV representing an immunodominant CTL epitope of luciferase (LucP; ref. ) in mice immunized with Luc DNA (pVaxLuc). The average number of IFN-γ producing splenocytes per mln cells in mice receiving pVaxLuc followed by electroporation using Dermavax (n = 6), or BEX devices at 100 V (n = 6), or empty vector pVax1 with electroporation using BEX device at 100 V (n = 6) after priming (day 21), boosting (day 30), and after challenge with 4T1luc2 cells (day 9, or 43 from the start) (see Methods for details) ( A ); FACS of the pooled splenocytes of Luc-gene immunized mice detecting LucP-specific secretion of IFN-γ by CD8 + T cells ( B ); Average LucP-specific IFN-γ response of mice immunized with Luc DNA and electroporated using CUY21EditII (BEX) at 50 V and 100 V ( C ). Red line indicates the cut-off for specific IFN-γ response as number of spots registered in non-immune (control and vector immunized) mice + 3 STDV. *p < 0,05, Luc DNA immunized mice electroporated at 100 V, and **p < 0,1, Luc DNA immunized mice electroporated at 50 V compared to vector immunized mice (Mann-Whitney test; STATISTICA AXA 10.0).

Article Snippet: 4T1 (CRL-2539TM, ATCC) and 4T1luc2 (BW124087, Perkin Elmer, Waltham, MA) were purchased; 4T1luc-D6 clone was described previously .

Techniques: Luciferase, Electroporation, Plasmid Preparation, Control, MANN-WHITNEY

Journal: Journal of Cellular and Molecular Medicine

Article Title: Obesity and triple‐negative‐breast‐cancer: Is apelin a new key target?

doi: 10.1111/jcmm.15639

Figure Lengend Snippet: Infusion of obesity‐related levels of apelin favours TNBC brain metastatization. A, Ex vivo bioluminescent signals from 4T1 Luc‐GFP in lungs of PBS or apelin‐infused (0.1 µmol/kg/d) Balb/c nude mice. B, Quantification of bioluminescent signals of lung metastases by intensity. C, Ex vivo bioluminescent signals from 4T1 Luc‐GFP in brains of PBS or Apelin‐infused Balb/c nude mice. D, Quantification of metastatic brains in PBS or Apelin‐infused mice. Number of mice per group for (A, B): Control: 9, Apelin: 9, and for (C, D): Control: 9, Apelin: 10. Data were analysed using two‐way ANOVA followed by Bonferroni post hoc test for (B). Data were analysed using Fisher's exact test for (D)

Article Snippet: The 4T1 Luc‐GFP cell line was obtained from Perkin Elmer (4T1 Red FLuc‐GFP, PerkinElmer, Waltham, MA, USA) and grows only in immunodeficient mice.

Techniques: Ex Vivo

Journal: Bioactive Materials

Article Title: Leukocyte/platelet hybrid membrane-camouflaged dendritic large pore mesoporous silica nanoparticles co-loaded with photo/chemotherapeutic agents for triple negative breast cancer combination treatment

doi: 10.1016/j.bioactmat.2021.04.004

Figure Lengend Snippet: Schematic illustrations for preparation method of LPHM@DDI NPs (A) and their synergistic effects in TNBC treatment via combining PTT/PDT with chemotherapy (B).

Article Snippet: Mouse TNBC cell line 4T1 and mouse macrophage cell line Raw 264.7 were sourced from BioVector NTCC (Beijing, China).

Techniques:

Journal: Bioactive Materials

Article Title: Leukocyte/platelet hybrid membrane-camouflaged dendritic large pore mesoporous silica nanoparticles co-loaded with photo/chemotherapeutic agents for triple negative breast cancer combination treatment

doi: 10.1016/j.bioactmat.2021.04.004

Figure Lengend Snippet: In vitro PTT/PDT performances and drug releases of LPHM@DDI NPs. IR thermal images of DLMSNs, DOX, IR780, DDI NPs, and LPHM@DDI NPs (A) and their temperature changes (B) within the 10-min laser irradiation period. (C) Fluorescence intensities of SOSG in the dispersions of DLMSNs, DOX, IR780, DDI NPs, and LPHM@DDI NPs after 2, 4, 6, 8, and 10 min of laser irradiation. (D) Fluorescence intensities of DCFH-DA in 4T1 cells after incubation of IR780, DOX, DDI NPs, and LPHM@DDI NPs alone (‒L) and followed by laser irradiation (+L). ** represents P < 0.01 vs. control; ## represents P < 0.01 between two groups. (E) In vitro release profiles of DOX from LPHM@DDI NPs alone and (F) their pre-procession with 10-min laser irradiation at different pH values.

Article Snippet: Mouse TNBC cell line 4T1 and mouse macrophage cell line Raw 264.7 were sourced from BioVector NTCC (Beijing, China).

Techniques: In Vitro, Irradiation, Fluorescence, Incubation, Control

Journal: Bioactive Materials

Article Title: Leukocyte/platelet hybrid membrane-camouflaged dendritic large pore mesoporous silica nanoparticles co-loaded with photo/chemotherapeutic agents for triple negative breast cancer combination treatment

doi: 10.1016/j.bioactmat.2021.04.004

Figure Lengend Snippet: Cellular uptake and intracellular localization of LPHM@DDI NPs. (A) Confocal microscopic images of 4T1 cells after 2 h of incubation with DDI, LM@DDI, PM@DDI, and LPHM@DDI NPs. Mean fluorescence intensities (MFIs) of DOX (B) and IR780 (C) in 4T1 cells after incubation with the above NPs. Flow cytometry analysis of Raw 264.7 cells after 1 h of incubation of DDI, LM@DDI, PM@DDI and LPHM@DDI NPs (D) and MFIs of intracellular DOX (E). ** represents P < 0.01 vs. control.

Article Snippet: Mouse TNBC cell line 4T1 and mouse macrophage cell line Raw 264.7 were sourced from BioVector NTCC (Beijing, China).

Techniques: Incubation, Fluorescence, Flow Cytometry, Control

Journal: Bioactive Materials

Article Title: Leukocyte/platelet hybrid membrane-camouflaged dendritic large pore mesoporous silica nanoparticles co-loaded with photo/chemotherapeutic agents for triple negative breast cancer combination treatment

doi: 10.1016/j.bioactmat.2021.04.004

Figure Lengend Snippet: In vitro synergistic anti-TNBC effects of LHPM@DDI NP-mediated combination treatment. Cytotoxicities of DOX with and without laser irradiation (A), IR780 with and without laser irradiation (B), and DDI NPs with and without laser irradiation (C) in 4T1 cells at different drug concentrations. (D) Cytotoxicities of DDI, LM@DDI, PM@DDI, and LPHM@DDI NPs with and without laser irradiation at the DOX and IR780 concentrations of 0.75 and 1.0 μg/mL, respectively. * and ** represent P < 0.05 and P < 0.01 between two groups. (E) Fluorescence microscopic images of 4T1 cells treated with LPHM@DDI NPs alone and followed by laser irradiation after co-staining of calcein-AM and ethidium homodimer-1. Green and red fluorescence indicate liver and dead cells. (F) Flow cytometry analysis of apoptosis in 4T1 cells treated with DDI, LM@DDI, PM@DDI, and LPHM@DDI NPs alone and followed by laser irradiation. The DOX and IR780 concentrations were 0.45 and 0.6 μg/mL, respectively. Four quadrants (Q1, Q2, Q3 and Q4) represent dead cells, late apoptotic cells, early apoptotic cells and live cells, respectively.

Article Snippet: Mouse TNBC cell line 4T1 and mouse macrophage cell line Raw 264.7 were sourced from BioVector NTCC (Beijing, China).

Techniques: In Vitro, Irradiation, Fluorescence, Staining, Flow Cytometry

Journal: Bioactive Materials

Article Title: Leukocyte/platelet hybrid membrane-camouflaged dendritic large pore mesoporous silica nanoparticles co-loaded with photo/chemotherapeutic agents for triple negative breast cancer combination treatment

doi: 10.1016/j.bioactmat.2021.04.004

Figure Lengend Snippet: Biodistribution and tumor accumulation of LPHM@DDI NPs in TNBC mice. (A) In vivo fluorescence images of 4T1 tumor-bearing mice at 6, 12, 24 and 48 h post-administration of PBS (the control), DDI NPs, LM@DDI NPs, PM@DDI NPs, and LPHM@DDI NPs. The dotted-red circle represents the tumor location in the control mouse. Ex vivo fluorescence images (B) and mean radiant efficiencies (MREs) (C) of the major organs and tumors isolated from the above mice. H, Li, S, Lu, K and T indicate the heart, liver, spleen, lung, kidney and tumor, respectively. * and ** represent P < 0.05 and P < 0.01 between two groups.

Article Snippet: Mouse TNBC cell line 4T1 and mouse macrophage cell line Raw 264.7 were sourced from BioVector NTCC (Beijing, China).

Techniques: In Vivo, Fluorescence, Control, Ex Vivo, Isolation

Journal: Bioactive Materials

Article Title: Leukocyte/platelet hybrid membrane-camouflaged dendritic large pore mesoporous silica nanoparticles co-loaded with photo/chemotherapeutic agents for triple negative breast cancer combination treatment

doi: 10.1016/j.bioactmat.2021.04.004

Figure Lengend Snippet: In vivo synergistic anti-TNBC effects of LPHM@DDI NP-mediated combination treatment. Tumor growth curves of 4T1 tumor-bearing mice treated with PBS (the control), the DOX/IR780 mixture, DDI NPs, LM@DDI NPs, PM@DDI, and LPHM@DDI NPs alone (A) and followed by laser irradiation (B). Photos of representative tumors sourced from the mice with various treatments (C) and microscopic images of their sections with H&E staining (D). (E) Photos of representative mice at 0, 5 and 18 d during the treatment period. Weight change curves of the mice receiving the above-mentioned treatments (F) and followed by laser irradiation (G). Microscopic images of tumor sections with TUNEL staining (H) and CD31 immunohistochemical staining (I) from the mice at 2 d during treatment period. The red arrows represent apoptotic cells or CD31-stained areas in tumor sections. (J) Comparison of MVDs calculated from CD31-stained tumor sections in various treatment groups. ** represents P < 0.01 vs. control; ## represents P < 0.01 between two groups.

Article Snippet: Mouse TNBC cell line 4T1 and mouse macrophage cell line Raw 264.7 were sourced from BioVector NTCC (Beijing, China).

Techniques: In Vivo, Control, Irradiation, Staining, TUNEL Assay, Immunohistochemical staining, Comparison