4t1 breast cancer murine cell line  (ATCC)

Journal: Contrast Media & Molecular Imaging

Article Title: 99m Tc-Radiolabeled TPGS Nanomicelles Outperform 99m Tc-Sestamibi as Breast Cancer Imaging Agent

doi: 10.1155/2019/4087895

Figure Lengend Snippet: (a) 99m Tc-sestamibi and 99m Tc-radiolabeled TPGS micelles cellular uptake (4T1 cell line). Results are expressed as the percentage of activity accumulated in the cellular fraction normalized by 10 6 cells. (b) 99m Tc-sestamibi and 99m Tc-radiolabeled TPGS micelles biodistribution in a breast cancer animal model developed with 4T1 murine cell line in BALB/c mice. Results are expressed as the percentage of injected dose in each organ normalized by organ weight (%ID/g). Bars represent mean ± SD. (c) Gamma camera imaging of 99m Tc-radiolabeled TPGS micelles (12 h after injection) and 99m Tc-sestamibi (15 min after injection) in a breast cancer animal model developed with 4T1 murine cell line in BALB/c mice. Green arrows indicate the palpable tumoral site. Representative images are shown. Mice outline was drawn for anatomical references.

Article Snippet: 4T1 Breast cancer murine cell line was obtained from American Type Culture Collection (ATCC® CRL-2539™ BALB/cfC3H strain).

Techniques: Activity Assay, Animal Model, Injection, Imaging

Journal: Nutrients

Article Title: A Diet Lacking Selenium, but Not Zinc, Copper or Manganese, Induces Anticancer Activity in Mice with Metastatic Cancers

doi: 10.3390/nu16142249

Figure Lengend Snippet: In vivo metastatic cancer models.

Article Snippet: CT26.WT (murine colorectal cancer cells, CRL-2638) and 4T1 (murine triple-negative breast cancer cells, CRL-2539), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vivo, Positive Control

Journal: Nutrients

Article Title: A Diet Lacking Selenium, but Not Zinc, Copper or Manganese, Induces Anticancer Activity in Mice with Metastatic Cancers

doi: 10.3390/nu16142249

Figure Lengend Snippet: The anticancer activity of cisplatin and a diet lacking selenium in mice with metastatic triple-negative breast cancer (4T1 murine breast cancer cells inoculated in the tail vein of female BALB/cAnNRj mice). ( a ) The survival of mice treated with cisplatin (intraperitoneal administration of 5 mg/kg once a week for 4 weeks), mice fed a control diet prepared from scratch, and a diet lacking selenium. ( b ) The body weights of the mice relative to their body weights at the beginning of treatments (day 4). Control: control diet prepared from scratch; C-Se: diet without selenium.

Article Snippet: CT26.WT (murine colorectal cancer cells, CRL-2638) and 4T1 (murine triple-negative breast cancer cells, CRL-2539), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: GCH1 induces immunosuppression through metabolic reprogramming and IDO1 upregulation in triple-negative breast cancer

doi: 10.1136/jitc-2021-002383

Figure Lengend Snippet: GTP cyclohydrolase 1 (GCH1) positively correlates with regulatory T cells (Tregs) infiltration in triple-negative breast cancer (TNBC). (A) Schematic diagram depicting the screening of metabolic genes that are positively related to Tregs infiltration in TNBC cohorts. (B) The protein and mRNA abundance of GCH1 in different subtypes of breast cancer tissues (n=9 each, Oslo2 Landscape cohort). Her2, human epidermal growth factor receptor 2; LumA, luminal A; LumB, luminal B. (C and D) Correlation between the GCH1 mRNA level and Tregs infiltration in the Fudan University Shanghai Cancer Center (FUSCC) cohort (C) and the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) TNBC cohort (D) determined by Pearson correlation analysis. (E) The infiltration of Tregs with GCH1 amplification in the METABRIC TNBC cohort. (F) GCH1 mRNA level in the adjacent normal and tumor tissues in the FUSCC cohort. (G) Immunohistochemistry detection of GCH1 in the FUSCC cohort. (H) Kaplan-Meier analysis of overall survival and disease-free survival in the FUSCC cohort (n=168 in the GCH1-low cohort and n=76 in the GCH1-high cohort). (I) Volumes of tumors harvested from mice injected with a total of 5×10 5 4T1 cells expressing control or Gch1 short hairpin RNA. (J) Representative dot plot of Tregs analyzed by flow cytometry (gated on CD4 + T cells) and the corresponding quantitative results (n=4 each). The data are presented as the mean±SD (E, I, and J): two-tailed unpaired Student’s t-test for E and J, paired t-test for F, and two-way analysis of variance (ANOVA) test for I. ** P <0.01 and *** P <0.001. FOXP3, forkhead box protein P3; shCtrl, short hairpin control; shGch1, short hairpin Gch1.

Article Snippet: The human breast cancer cell lines BT549, Hs578T, and MDA-MB-453 and human embryonic kidney (HEK) 293 T cells, together with the murine breast cancer cell line 4T1, were obtained from American Type Culture Collection and maintained under standard conditions.

Techniques: Amplification, Immunohistochemistry, Injection, Expressing, shRNA, Flow Cytometry, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: GCH1 induces immunosuppression through metabolic reprogramming and IDO1 upregulation in triple-negative breast cancer

doi: 10.1136/jitc-2021-002383

Figure Lengend Snippet: GTP cyclohydrolase 1 (GCH1) increases regulatory T cells (Tregs) infiltration, reduces apoptosis, and increases the programmed cell death-1 (PD-1) + fraction. Tumors were harvested from mice with orthotopic injection of a total of 3×10 5 4T1 cells expressing a control empty vector (Vec) or GCH1 (n=6 each). (A) Tumor volumes. (B) Representative dot plot of Tregs analyzed by flow cytometry and the corresponding quantitative results. (C) Representative dot plot of Tregs apoptosis analyzed by flow cytometry and the corresponding quantitative results. (D) Representative histograms of PD-1 on Tregs analyzed by flow cytometry and the corresponding quantitative results. (E, F, G, and H) Quantitative results of CD8 + T cell infiltration (E), apoptosis (F), PD-1 expression (G), and interferon-γ (IFN-γ) production (H) analyzed by flow cytometry. (I) Volumes of tumors from mice with orthotopic injection of a total of 3×10 5 4T1 cells expressing Vec or GCH1. The mice were treated with isotype IgG or an anti-CD25 antibody after the tumors were palpable. (J and K) Representative dot plot or histograms and the corresponding quantitative results of apoptosis (J) and PD-1 expression (K) of Tregs cocultured with BT549 cells expressing Vec or GCH1. The data are presented as the mean±SD (A, B, C, D, E, F, G, H, I, J, and K). Two-way analysis of variance (ANOVA) test for A and I; two-tailed unpaired Student’s t-test for B, C, D, E, F, G, H, J, and K; n=3 independent experiments (J and K). * P <0.05; ** P <0.01; *** P <0.001; n.s., not significant. FOXP3, forkhead box protein P3; iso, isotype; SSC, side scatter.

Article Snippet: The human breast cancer cell lines BT549, Hs578T, and MDA-MB-453 and human embryonic kidney (HEK) 293 T cells, together with the murine breast cancer cell line 4T1, were obtained from American Type Culture Collection and maintained under standard conditions.

Techniques: Injection, Expressing, Plasmid Preparation, Flow Cytometry, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: GCH1 induces immunosuppression through metabolic reprogramming and IDO1 upregulation in triple-negative breast cancer

doi: 10.1136/jitc-2021-002383

Figure Lengend Snippet: GTP cyclohydrolase 1 (GCH1) upregulates indoleamine 2,3-dioxygenase 1 (IDO1) via aryl hydrocarbon receptor (AHR) activation. (A) Gene set enrichment analysis plot showing enriched AHR signature in the Fudan University Shanghai Cancer Center (FUSCC), Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) triple-negative breast cancer (TNBC), and The Cancer Genome Atlas (TCGA) TNBC cohorts. (B) Representative immunofluorescence image of AHR in BT549 cells expressing vector (Vec) or GCH1. Scale bar=10 µm. (C) Cytoplasmic and nuclear protein fractions of the AHR protein in cells expressing Vec or GCH1. (D) Relative IDO1 transcription levels detected by quantitative reverse transcription PCR (qRT-PCR) analysis and IDO1 levels detected by western blot in cells expressing Vec or AHR. (E) Relative IDO1 and AHR transcription levels detected by qRT-PCR analysis and IDO1 levels detected by western blot in BT549 cells expressing Vec or GCH1 transfected with small interfering RNA targeting AHR (si AHR) . (F) Relative Ido1 and Ahr transcription levels detected by qRT-PCR analysis and IDO1 levels detected by western blot in 4T1 cells expressing Vec or GCH1 treated with si Ahr . The data are presented as the mean±SD (D, E, and F): two-tailed unpaired Student’s t-test for D, E, and F; n=3 independent experiments (D, E and F). *** P <0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HDAC1, histonedeacetylase 1; NES, normalized enrichment score; siAHR, small interfering RNA targeting AHR.

Article Snippet: The human breast cancer cell lines BT549, Hs578T, and MDA-MB-453 and human embryonic kidney (HEK) 293 T cells, together with the murine breast cancer cell line 4T1, were obtained from American Type Culture Collection and maintained under standard conditions.

Techniques: Activation Assay, Immunofluorescence, Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Transfection, Small Interfering RNA, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: GCH1 induces immunosuppression through metabolic reprogramming and IDO1 upregulation in triple-negative breast cancer

doi: 10.1136/jitc-2021-002383

Figure Lengend Snippet: 2,4-Diamino-6-hydroxypyrimidine (DAHP) reverses the immune suppression induced by GTP cyclohydrolase 1 (GCH1). (A) Tetrahydrobiopterin (BH4) levels detected with ELISA in the supernatants of BT549 cells expressing vector (Vec), GCH1, or GCH1 mutations (A184H and M211I). (B and C) Indoleamine 2,3-dioxygenase 1 (IDO1) levels (B) and the cytoplasmic and nuclear protein fractions of the aryl hydrocarbon receptor (AHR) protein (C) detected by western blot in BT549 cells expressing Vec, GCH1, or GCH1 mutations (A184H and M211I). (D) Apoptosis and programmed cell death-1 (PD-1) levels of regulatory T cell (Tregs) cocultured with BT549 cells expressing Vec, GCH1, or GCH1 mutations (A184H and M211I) and analyzed by flow cytometry. (E) IDO1 levels in MDA-MB-453 cells treated with different concentrations of DAHP for 24 hours determined by western blot analysis. (F) Tumor volumes. Tumors were harvested from mice with orthotopic injection of a total of 3×10 5 4T1 cells overexpressing GCH1. Mice were treated with DAHP, an anti-PD-1 antibody, or the combination after the tumors were palpable (n=6 each). (G) Quantitative results of infiltration and apoptosis in Tregs and infiltration, apoptosis, and interferon-γ (IFN-γ) production in CD8 + T cells analyzed by flow cytometry. (H) Schematic diagram of GCH1 inducing immunosuppression in triple-negative breast cancer. GCH1 reprogrammed tryptophan (Trp) metabolism, increased the intracellular level of L-5-hydroxytryptophan (5-HTP), activated AHR, upregulated IDO1, and finally induced an immunosuppressive environment. DAHP could reverse the GCH1-induced immunosuppression and enhance the antitumor effect of PD-1 blockade immunotherapy. The data are presented as the mean±SD (A, D, F, and G): two-tailed unpaired Student’s t-test for A, D, and G; two-way analysis of variance (ANOVA) test for F; and n=3 independent experiments (A and D). * P <0.05; ** P <0.01; *** P <0.001; n.s., not significant. DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HDAC1, histonedeacetylase 1; iso, isotype; Kyn, kynurenine.

Article Snippet: The human breast cancer cell lines BT549, Hs578T, and MDA-MB-453 and human embryonic kidney (HEK) 293 T cells, together with the murine breast cancer cell line 4T1, were obtained from American Type Culture Collection and maintained under standard conditions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Plasmid Preparation, Western Blot, Flow Cytometry, Injection, Two Tailed Test

Journal: Nanotheranostics

Article Title: Effects of molecular weight and structural conformation of multivalent-based elastin-like polypeptides on tumor accumulation and tissue biodistribution

doi: 10.7150/ntno.39804

Figure Lengend Snippet: In vivo biodistribution imaging of AP1-ELPs in allograft mice model. (A) FPR 675-labeled E60, A38, A60, A86, and A100 (100 µ M) were injected intravenously into 4T1 tumor allograft mice. The in vivo fluorescence images were collected at different time points (10 min, 1, 2, 4, 6, 12, 24, and 48 h) after intravenous injection (IV) to determine biodistribution and a representative image of each group is shown. (B) Quantification of fluorescence intensities in tumor sites at different time intervals. All data are presented as mean ± SD (n = 6), statistical significance was determined by one-way ANOVA analysis, (*** P <0.001, ** P <0.01). (C) Fluorescence intensities of excised tumors and organs at 48 h post-injection (n = 10). Data are represented as mean ± SD and analyzed using one-way ANOVA, (n=3), * P <0.05.

Article Snippet: 4T1 murine breast cancer cells and MDA MB231 human breast cancer cells were obtained from the American Type Culture Collection (ATCC).

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

Journal: Nanotheranostics

Article Title: Effects of molecular weight and structural conformation of multivalent-based elastin-like polypeptides on tumor accumulation and tissue biodistribution

doi: 10.7150/ntno.39804

Figure Lengend Snippet: In vivo biodistribution of AP1-ELPs in orthotopic mice model. (A) FPR 675-labeled E60, A38, A60, A86, and A100 (100 µ M) were injected intravenously into 4T1 tumor bearing orthotopic mice after five days of implantation. The representative IVIS images were collected at different time points were shown. (B) The fluorescence intensities in tumor sites was determined at different time points (0, 6, 12, 24, 48 h) respectively. The data presented as mean ± SEM (n=3), the statistically significance were calculated using one-way ANOVA with Bonferroni correction. ** P <0.01, * P <0.05 (C) Histogram represents the fluorescence intensities of excised tumors and organs after 48 h post-injection. All Data were represented as mean ± SD (n = 3) and analyzed by one-way ANOVA, * P <0.05.

Article Snippet: 4T1 murine breast cancer cells and MDA MB231 human breast cancer cells were obtained from the American Type Culture Collection (ATCC).

Techniques: In Vivo, Labeling, Injection, Fluorescence

Journal: Nanotheranostics

Article Title: Effects of molecular weight and structural conformation of multivalent-based elastin-like polypeptides on tumor accumulation and tissue biodistribution

doi: 10.7150/ntno.39804

Figure Lengend Snippet: Immunohistology staining of tumor tissues extracted from 4T1 tumor xenograft mice after 48 h post-intravenous injection of respective polypeptides. Tumor tissue sections were stained with anti-IL-4R and observed under a confocal microscope. Representative images of 10 repetitive experiments. Blue, nuclei stained with DAPI; Green, tumor cells stained with anti-IL-4R; Red, labeled polypeptide. Scale bar, 50 µm.

Article Snippet: 4T1 murine breast cancer cells and MDA MB231 human breast cancer cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Staining, Injection, Microscopy, Labeling

Journal: International Journal of Nanomedicine

Article Title: Docosahexaenoic acid liposomes for targeting chronic inflammatory diseases and cancer: an in vitro assessment

doi: 10.2147/IJN.S115995

Figure Lengend Snippet: Effect of docosahexaenoic acid-loaded liposomes (ω-liposomes) and control liposomes (C-liposomes) on tumor-cell proliferation. Notes: ( A ) FaDu cells and ( B ) 4T1 cells were exposed for 24 hours to ω-liposomes and C-liposomes, after which the medium was replaced with medium containing bromodeoxyuridine (BrdU) and cells incubated for additional 4–6 hours. Afterward, BrdU incorporation was determined by enzyme-linked immunosorbent assay. Data presented as mean ± standard error of mean from three independent experiments (each n$4). * P <0.05, *** P <0.001 compared to control nontreated (NT) cells (Student’s t -test). Abbreviation: TL, total lipid.

Article Snippet: Murine 4T1 breast cancer cells and human FaDu squamous cell carcinoma cells were obtained from the ATCC.

Techniques: Incubation, BrdU Incorporation Assay, Enzyme-linked Immunosorbent Assay