Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Therapeutic impact of Nintedanib with paclitaxel and/or a PD-L1 antibody in preclinical models of orthotopic primary or metastatic triple negative breast cancer

doi: 10.1186/s13046-018-0999-5

Figure Lengend Snippet: Paclitaxel and its combination with nintedanib increased median survival in the advanced metastatic breast cancer LM2–4 model. a ) Kaplan-Meier survival curves and median survival values. Paclitaxel (PTX) significantly increased median survival compared to the control group ( p = 0.033; Log-rank (Mantel Cox) Test, n = 8–10). Combination therapy increased median survival (81 days vs 60.5 days, control group) but it did not reach significance. Treatment started around 40 days after cell implantation. b) Effect of sunitinib alone and when combined with PTX in the advanced metastatic LM2–4 breast cancer model. Kaplan-Meier survival curves and median survival values. Modified from Guerin et al., 2013 . PTX alone increased survival whereas sunitinib alone did not, and adding sunitinib to PTX did not result in increased efficacy

Article Snippet: The EMT-6 (ATCC® CRL-2755TM) mouse breast cancer cell line and the derived variant EMT-6/CDDP -selected in vivo for acquired resistance to cisplatin [ ]-, were cultured in DMEM medium with 5% FBS at 37 °C in 5% CO 2.

Techniques: Control, Modification

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Therapeutic impact of Nintedanib with paclitaxel and/or a PD-L1 antibody in preclinical models of orthotopic primary or metastatic triple negative breast cancer

doi: 10.1186/s13046-018-0999-5

Figure Lengend Snippet: Improvement of immunotherapy efficacy when treating primary tumors with nintedanib combination therapy . a ) Tumor growth in the primary EMT-6 breast cancer model. Treatment was started when average tumor volume was 120mm 3 , around 7 days after cell implantation. Statistical analysis on day 27. ANOVA followed by Tukey’s Multiple Comparison Test * p < 0.05, ** p < 0.01. Data are presented as means ± SD, n = 6. Region of flat line in the curves means that tumor in remaining mice had regressed, and in the case of mice treated with PD-L1 antibody, tumors grew back. Mice were treated with nintedanib and/or paclitaxel (PTX) for 70 days, and then treatment stopped. b) Tumor growth in the primary EMT-6/CDDP model. Treatment was started when average tumor volume was 120mm 3 , 7 days after cell implantation. Statistical analysis on day 27. Kruskal-Wallis test followed by Dunn’s Multiple Comparison Test, ** p < 0.01. Data are presented as means ± SD, n = 9–12. c-f) Effect of nintedanib, paclitaxel, anti-PD-L1 and the drug combinations on c) Vascularity; d) Proliferation; e) CD8+ tumor infiltrating cells; and f) Level of Necrosis. Histology and immunohistochemistry analyses were performed on tumor samples obtained from mice implanted with EMT-6/CDDP cells. Treatment was started when average tumor volume was 120mm 3 and all mice were sacrificed after 10 days of treatment. The Mann-Whitney test was used for statistical analyses. Data are presented as means ± SD, n = 6–7

Article Snippet: The EMT-6 (ATCC® CRL-2755TM) mouse breast cancer cell line and the derived variant EMT-6/CDDP -selected in vivo for acquired resistance to cisplatin [ ]-, were cultured in DMEM medium with 5% FBS at 37 °C in 5% CO 2.

Techniques: Comparison, Immunohistochemistry, MANN-WHITNEY

Journal: NPJ Breast Cancer

Article Title: NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

doi: 10.1038/s41523-019-0106-x

Figure Lengend Snippet: NDRG4 depletion in breast tumor cells increases lymph node adhesion and cell migration toward VN. a NDRG4 expression meta-analysis in human breast cancer cell lines using GOBO application . Breast cancer subtypes can be identified by different colors: red = Basal A, gray = Basal B and blue = Luminal cells. b Analysis of NDRG4 mRNA expression levels in MCF-7 cells transfected with NDRG4-shRNAs or scramble control (shSCR). Expression levels are relative to wild type cells and normalized to hydroxymethylbilane synthase (HMBS) gene. Error bars represent SEM of biological replicates ( n = 5). *** p < 0.001, ns = not significant, by one-way ANOVA. c Western blot analysis of NDRG4 expression in cytoplasmic extracts of MCF-7 shNDRG4s or shSCRs cells, using antibodies against NDRG4 and Actin. MCF-7 cells express all the three isoforms of NDRG4: 37 kDa (NDRG4-B), 39 kDa (NDRG4-B var ) and 41 kDa isoform (NDRG4-H). For simplicity, results obtained for two independent shNDRG4 clones were grouped and presented as the shNDRG4 group. ( d , left) Representative images of adherent red fluorescent MCF-7 cells on frozen rat lymph node sections. d , e Quantification of adherent MCF7 ( d ) and T47D ( e ) breast tumor cells in lymph nodes sections (right, n = 4 independent experiments for MCF-7 and 3 for T47D, ** p < 0.01 by two-tailed t test). f Stationary adhesion assays showed that NDRG4 knockdown promotes ‘adhesive switch’ between FN and VN. Error bars represent SEM of biological replicates ( n = 4). Cells do not adhere to albumin control (BSA, data not shown). * p < 0.01, ** p < 0.01, ns = not significant by two-tailed t tests. g , h Haptotactic cell migration toward VN was analyzed by transwell migration assay in MCF-7 ( g ) and T47D ( h ) cell lines. Error bars represent SEM of biological replicates ( n = 4). *** p < 0.001 by two-tailed t tests

Article Snippet: For stable transfection of NDRG4 shRNA into MCF-7 and T47D breast tumor cells, the pGFP-V-RS vector containing shRNA inserts which targeted the 3′-unttranslated region of NDRG4 (TRCN0000134583 and TRCN0000137216) was purchased (Origene).

Techniques: Migration, Expressing, Transfection, Western Blot, Clone Assay, Two Tailed Test, Transwell Migration Assay

Journal: NPJ Breast Cancer

Article Title: NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

doi: 10.1038/s41523-019-0106-x

Figure Lengend Snippet: NDRG4 knockdown promotes clustering of β1-integrin at the leading edge of T47D cells. Representative confocal images of β1 integrin subunit (MAB1965, green) at the ventral cell surface of VN-adherent T47D shNDRG4 or shSCR cells

Article Snippet: For stable transfection of NDRG4 shRNA into MCF-7 and T47D breast tumor cells, the pGFP-V-RS vector containing shRNA inserts which targeted the 3′-unttranslated region of NDRG4 (TRCN0000134583 and TRCN0000137216) was purchased (Origene).

Techniques: