Journal: International journal of cancer

Article Title: TP53 Induced Glycolysis and Apoptosis Regulator and Monocarboxylate Transporter 4 drive metabolic reprogramming with c-MYC and NFkB activation in breast cancer

doi: 10.1002/ijc.34660

Figure Lengend Snippet: Effect of Lactate on carcinoma cell compartment. (A) MCT1 immunoblot in MCF7 cells under control conditions and after treatment with 5 mM lactate. (B) c-MYC immunoblot in MCF7 carcinoma cells exposed to 5 mM Lactate compared with control. (C) c-MYC immunoblot in MCF7 and MDA-MB-231 Sg Control or MCF7 sg TIGAR knockdown cells. (D) c-MYC and TIGAR immunoblot in MCF7 and AT-3 TIGAR overexpressing cells compared with EV empty vector control cells. (E) c-MYC and TIGAR immunoblot in MCF7 Sg MYC knockdown cells compared with Sg Control cells. (F) c-MYC immunoblot in AT-3 and HA-cMYC immunoblot PY8119 in c-MYC overexpressing cells compared with LV control cells. (G) AT-3 and PY8119 LV control and c-MYC overexpressing cells were stably transfected with a secrete-pair dual luminescence GLuc/SEAP TIGAR promoter reporter. The GLuc luminescence is produced based on TIGAR promoter luciferase activity. The GLuc luminescence is normalized to SEAP luminescence which allows for transfection and expression normalization (* P < .05). (H) AT-3 LV control and c-MYC overexpressing cells were stably transfected with a secrete-pair dual luminescence GLuc/SEAP mutant TIGAR promoter reporter. The GLuc luminescence is produced based on TIGAR promoter luciferase activity. The GLuc luminescence is normalized to SEAP luminescence which allows for transfection and expression normalization (* P < .05). (I) TIGAR, c-MYC and H3K9 acetylation immunoblot in MCF7 Sg Control and Sg TIGAR knockdown cells under control conditions or after exposure to 2.5 mM acetate. (J) Lactyllysine immunoblot in MCF7 Sg control and Sg TIGAR knockdown cells exposed to 0, 5 and 10 mM Lactate. Quantification of Lactyllysine protein expression normalized to vinculin in MCF7 control and TIGAR knockdown cells exposed to lactate. (K) Lactyllysine immunoblot in MCF7 EV empty vector control or TIGAR overexpressing cells exposed to 0, 5 and 10 mM Lactate. Quantification of Lactyllysine protein expression normalized to vinculin in MCF7 control and TIGAR overexpressing cells exposed to lactate. Error bars represent the SE of the mean.

Article Snippet: The PY8119 mouse mammary carcinoma (RRID: CVCL_AQ09) cell line was obtained from ATCC.

Techniques: Western Blot, Control, Knockdown, Plasmid Preparation, Stable Transfection, Transfection, Produced, Luciferase, Activity Assay, Expressing, Mutagenesis

Journal: International journal of cancer

Article Title: TP53 Induced Glycolysis and Apoptosis Regulator and Monocarboxylate Transporter 4 drive metabolic reprogramming with c-MYC and NFkB activation in breast cancer

doi: 10.1002/ijc.34660

Figure Lengend Snippet: Tumor growth and mitochondrial metabolism induced by TIGAR is mediated by MCT4 in fibroblasts. (A, B) Tumor volume and weight of EV empty vector control or TIGAR overexpressing breast carcinoma cells co-injected with wild-type (WT) or MCT4−/− MEFs into the flank of WT or MCT4−/− C57Bl6 mice. (C) Model of effects of TIGAR expression in carcinoma cells. TIGAR in carcinoma cells positively modulates the expression c-MYC in a cell autonomous fashion and modulates the expression of fibroblast MCT4 and CAV1. Conversely, fibroblast MCT4 and CAV1 expression modulates the expression of TIGAR in carcinoma cells. This cross-talk is mediated by the production of the soluble factors TGFB, IL6 and lactate. (D) Oxygen consumption rate (OCR) of tumors generated from PY8119 EV empty vector control or TIGAR overexpressing breast carcinoma cells co-injected with wild-type (WT) MEFs in WT mice was measured under baseline conditions with 5 mM glucose, after injection of 1.5 μM Oligomycin and after injection of 2.0 μM FCCP (N = 6). (E) Oxygen consumption rate (OCR) of tumors generated from PY8119 EV empty vector control or TIGAR overexpressing breast carcinoma cells co-injected with wild-type (WT) MEFs in WT mice was measured under baseline conditions with 5 mM glucose and 10 mM lactate, after injection of 1.5 μM Oligomycin and after injection of 2.0 μM FCCP (N = 6). (F) Oxygen consumption of tumors generated from PY8119 EV empty vector control or TIGAR overexpressing breast carcinoma cells co-injected with MCT4−/− MEFs in MCT4−/− mice was measured under baseline conditions with 5 mM glucose, after injection of 1.5 μM Oligomycin, and after injection of 2.0 μM FCCP (N = 6). G, Oxygen consumption of tumors generated from PY8119 EV empty vector control or TIGAR overexpressing breast carcinoma cells co-injected with MCT4−/− MEFs in MCT4−/− mice was measured under baseline conditions with 5 mM glucose and 10 mM lactate, after injection of 1.5 μM Oligomycin, and after injection of 2.0 μM FCCP (N = 6). OCR values were normalized to protein concentration. Error bars represent the SE of the mean. (* P < .05). Created with BioRender.com

Article Snippet: The PY8119 mouse mammary carcinoma (RRID: CVCL_AQ09) cell line was obtained from ATCC.

Techniques: Plasmid Preparation, Control, Injection, Expressing, Generated, Protein Concentration

Journal: Breast Cancer Research : BCR

Article Title: Secreted frizzled-related protein 2 monoclonal antibody-mediated IFN-ϒ reprograms tumor-associated macrophages to suppress triple negative breast cancer

doi: 10.1186/s13058-025-02176-6

Figure Lengend Snippet: SFRP2 mAb treatment reduces the number of metastases and is associated with an increase in apoptosis in tumors, as well as an increase in the M1/M2 TAM ratio and in the levels of IFN-ƴ. A EO771.LMB tumor cells were injected via tail vein into C57/BL6 mice and treated with IgG1 control for 7 days (representative lung picture) and hSFRP2 mAb 8 mg/kg iv q for 3 days (representative lung picture). After 3 weeks, the lungs were resected, and surface metastatic lesions were counted. The bar graph indicates a higher incidence of lung metastases in IgG1 control-treated mice (gray, n = 15) than in hSFPR2 mAb-treated mice (black, n = 15, p < 0.05). B TUNEL assay on FFPE EO771.LMB lung sections from mice treated with IgG1 control and hSFRP2 mAb show significantly more apoptotic cells in the hSFRP2 mAb treatment group (n = 15, * p , < 0.05) C PY8119 cells were injected via tail vein into C57/BL6 mice and treated with IgG1 control (n = 11) versus hSFRP2 mAb (n = 11). The bar chart shows a statistically significant reduction in lung metastases in hSFRP2 mAb-treated mice compared to the control ( p < 0.05). D FFPE PY8119 lung sections were stained with TUNEL assay, and the number of apoptotic cells/20 × HPF were counted. There was a significant increase in apoptotic cells in lung metastases from mice treated with hSFRP2 mAb (n = 10) compared to IgG1 control (n = 10, * p < 0.05). ( E ) FFPE E0771 lung sections were stained with antibodies to CD86, CD163, and F4/80. M1/M2 ratios for hSFRP2 mAb treated mice were increased compared to IgG1 treated mice (* p = 0.028, n = 3). F FFPE PY8119 lung sections were stained with antibodies against CD86, CD163, and F4/80. M1/M2 ratios for hSFRP2 mAb treated mice were increased compared to IgG1 treated mice (* p = 0.036, n = 3). ( G ) ELISA revealing an increase of IFN-ϒ protein in the serum of EO771.LMB ( p < 0.0001 n = 5) and PY8119 ( p < 0.0001 n = 7) tumor-bearing mice following treatment with hSFRP2 mAb

Article Snippet: Ten serum samples from EO771.LMB tumor-bearing mice (5 IgG1, 5 hSFRP2 treated) and 14 samples from PY8119 tumor-bearing mice (7 IgG1, 7 hSFRP2 treated) were analyzed for the presence of IFN-Υ using ELISA (R&D Systems, Minneapolis, MN, cat # MIF00) according to the manufacturer’s protocol.

Techniques: Injection, Control, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay