Journal: Cell Death & Disease

Article Title: Targeting ENO1 reprograms macrophage polarization to trigger antitumor immunity and improves the therapeutic effect of radiotherapy

doi: 10.1038/s41419-026-08416-7

Figure Lengend Snippet: A A total of 2.5 × 10 5 CT26 cells were subcutaneously injected into BALB/c mice for 11 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 20 mg/kg) five times at three-day intervals ( n = 5). The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 5). B The resected tumors from the CT26 tumor-bearing mice were analyzed by immunohistochemical analysis. ** p < 0.01. Unpaired t test ( n = 5). C A total of 2.5 × 10 5 CT26 cells were intravenously injected into BALB/c mice for 11 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 40 mg/kg) five times at three-day intervals ( n = 5). The tumor volume was recorded every three days. The resected lung tissues were analyzed by HE staining. *** p < 0.001. Unpaired t test ( n = 5). D 4T1 cells (5 × 105) were intravenously injected into BALB/c mice, which were then intraperitoneally administered anti-ENO1 antibodies (HuL001, 40 mg/kg) on the indicated days (n = 5). The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 5). E The resected tumors from 4T1-bearing mice were analyzed by HE staining. ** p < 0.01. Unpaired t test ( n = 5). F The resected lung tissues from 4T1-bearing mice were analyzed by HE staining, immunofluorescence staining and immunohistochemical analysis. ** p < 0.01 and *** p < 0.001. Unpaired t test ( n = 5). G The resected liver tissues from 4T1-bearing mice were analyzed by HE staining, immunofluorescence staining and immunohistochemical analysis. *** p < 0.001. Unpaired t test ( n = 5). H The density of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors from CT26- and 4T1-bearing mice was evaluated by immunofluorescence staining ( n = 3). I The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors from CT26-bearing mice. * p < 0.05 and *** p < 0.001. Unpaired t test ( n = 5). J The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors from 4T1-bearing mice. * p < 0.05 and *** p < 0.001. Unpaired t test ( n = 5). K The DEGs between HuL001- and Veh-treated resected CT26 tumors are shown in a volcano plot ( n = 2). L Compared with those in Veh.-treated resected tumors, the number of glycolysis-related gene sets significantly decreased in HuL001-treated resected tumors ( n = 2).

Article Snippet: The mouse colon cancer cell line CT26, mouse breast cancer cell line 4T1, human CRC cell lines (SW480, SW620, HCT116, LoVo and HT29) and human breast cancer cell lines (MDA-MB-468, BT-20, HS578T and SUM-159) were obtained from the ATCC.

Techniques: Injection, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Cell Death & Disease

Article Title: Targeting ENO1 reprograms macrophage polarization to trigger antitumor immunity and improves the therapeutic effect of radiotherapy

doi: 10.1038/s41419-026-08416-7

Figure Lengend Snippet: A A total of 5 × 10 5 CT26 cells were subcutaneously injected into the left legs of BALB/c mice for 5 days and then intraperitoneally administered with anti-ENO1 antibodies (HuL001, 40 mg/kg) or clodronate liposomes (50 μL/mouse) on the indicated days ( n = 5). Local radiotherapy was given on Day 10. The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 4). B The densities of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors were analyzed by immunofluorescence staining. The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors. * p < 0.05 and *** p < 0.001. One-Way ANOVA test ( n = 3). C 4T1 cells (5 ×10 4 ) were subcutaneously injected into the left legs of BALB/c mice for 4 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 20 mg/kg) six times on the indicated days ( n = 5). Local radiotherapy was given on Days 10 and 12. The tumor volume was recorded every three days. * p < 0.05 and *** p < 0.001. Two-Way ANOVA test ( n = 4). D The resected tumors were weighed on Day 40. * p < 0.05. One-Way ANOVA test ( n = 4). E The densities of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors were analyzed by immunofluorescence staining. F The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors. * p < 0.05. One-Way ANOVA test ( n = 3). G The frequencies of M1 (CD11c + CD11b + F4/80 + CD45 + 7AAD - CD3 - CD19 - ) and M2 (CD1206 + CD11b + F4/80 + CD45 + 7AAD - CD3 - CD19 - ) tumor-infiltrating macrophages were analyzed by flow cytometry. * p < 0.05. One-Way ANOVA test ( n = 3-4). H The quantification of the M1/M2 ratio is shown. * p < 0.05. One-Way ANOVA test ( n = 3). I A representative image of flow cytometric analysis of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells. J The frequency of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells was analyzed by flow cytometry. * p < 0.05. One-Way ANOVA test ( n = 3-4). K The density of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells is shown. * p < 0.05. One-Way ANOVA test ( n = 3-4). L The density of IFNγ + (IFNγ hi CD8 + CD3 + CD45 + 7AAD - ) T cells is shown ( n = 3-4). * p < 0.05 and ** p < 0.01. One-Way ANOVA test ( n = 3-4). M The proposed mechanism of TGFβ1/TGFβR/Smad3/PRMT5-mediated ENO1 translocation for lactate release via MCT4.

Article Snippet: The mouse colon cancer cell line CT26, mouse breast cancer cell line 4T1, human CRC cell lines (SW480, SW620, HCT116, LoVo and HT29) and human breast cancer cell lines (MDA-MB-468, BT-20, HS578T and SUM-159) were obtained from the ATCC.

Techniques: Injection, Liposomes, Immunofluorescence, Staining, Flow Cytometry, Translocation Assay

Journal: Oncogene

Article Title: TBKBP1 induces capecitabine resistance through negative regulation of type I interferon pathway in triple-negative breast cancer

doi: 10.1038/s41388-025-03598-4

Figure Lengend Snippet: A Schematic outline of Tbkbp1-knockdown in tumors induced by capecitabine: 4T1 mouse breast cancer cells stably expressing shNC or shTbkbp1 were subcutaneously injected into BALB/c mice. The mice were treated with capecitabine (25 mg/kg i.g.), or PBS every two days since Day 9. ( n = 8 mice/group). B Representative images of the tumors illustrating the effect of TBKBP1-depletion in the PBS and CAP groups. ( n = 8 mice/group). C Effects of Tbkbp1 knockdown on tumor growth in the PBS and CAP groups. The data were represented as the mean ± SEM. The two-way ANOVA was used to compare the data. D Tumor weights of the six groups. The data were represented as the mean ± SEM. Data were compared by using the Student’s t test. E Bar plots showing the percentages of CD3e + cells among CD45 + cells, CD4 + cells among CD3e + CD45 + cells, CD8 + cells among CD3e + CD45 + cells, interferon-γ (IFNγ)-positive cells among CD8 + T cells, CD86 + cells among CD45 + CD11b + F4/80 + cells, and CD206 + cells among CD45 + CD11b + F4/80 + cells. Data were compared by using the Student’s t test. F Representative IHC images of CD4, CD8, CD86, and CD206 expression in shNC and shTbkbp1 tumors in the PBS and capecitabine group. Scale bar, 20 μm. G Boxplots showing the percentages of CD4-, CD8-, CD86-, and CD206-positive cells. The data were represented as the mean ± SEM. Data were compared by using the Student’s t test.

Article Snippet: The human embryonic kidney cell line HEK293T, human TNBC cell lines Hs 578 T, MDA-MB-468, BT-549, SUM159, LM2-4175, HCC1806, and the mouse TNBC cell line 4T1 were obtained from American Type Culture Collection (ATCC).

Techniques: Knockdown, Stable Transfection, Expressing, Injection

Journal: Oncogene

Article Title: TBKBP1 induces capecitabine resistance through negative regulation of type I interferon pathway in triple-negative breast cancer

doi: 10.1038/s41388-025-03598-4

Figure Lengend Snippet: A Bar plots of GO and KEGG analysis results showing that Tbkbp1 knockdown in CAP-treated breast tumors upregulated several pathways and biological processes. B GSEA analysis revealed that Tbkbp1 knockdown promoted CAP-induced upregulation of genes involved in interferon-related pathways. C Heatmaps showing that Tbkbp1-depletion upregulated genes involved in IFN- α pathway (left) and IFN- γ pathway (right). D qRT-PCR results showing that knockdown of Tbkbp1 upregulated the mRNA levels of Tbk1, Irf3, and Ifngr1 in 4T1 cells (upper) and 4T07 cells (bottom) under 5-FU treatment. The data were represented as the mean ± SEM. Data were compared by using the Student’s t test. E qRT-PCR results showing that overexpression of Tbkbp1 downregulated the mRNA levels of Irf3 and Ifngr1 in AT3 cells (upper) and TS/A cells (bottom) under 5-FU treatment. The data were represented as the mean ± SEM. Data were compared by using the Student’s t-test. F Western blot analysis showing that Tbkbp1 knockdown increased the protein levels of Tbk1, Irf3, and Ifngr1 in 4T1 cells (left) and 4T07 cells (right) induced by 5-FU. Densitometric quantification of Western blots was performed using ImageJ software. G Western blot analysis revealed that Tbkbp1 overexpression decreased the protein levels of Tbk1, Irf3, and Ifngr1 in AT3 cells (left) and TS/A cells (right) induced by 5-FU. Densitometric quantification of Western blots was performed using ImageJ software.

Article Snippet: The human embryonic kidney cell line HEK293T, human TNBC cell lines Hs 578 T, MDA-MB-468, BT-549, SUM159, LM2-4175, HCC1806, and the mouse TNBC cell line 4T1 were obtained from American Type Culture Collection (ATCC).

Techniques: Knockdown, Quantitative RT-PCR, Over Expression, Western Blot, Software

Journal: Oncogene

Article Title: TBKBP1 induces capecitabine resistance through negative regulation of type I interferon pathway in triple-negative breast cancer

doi: 10.1038/s41388-025-03598-4

Figure Lengend Snippet: A Bar plots of GSEA, GO, KEGG, and Reactome analysis results based on RNA-seq data of breast tumors from BALB/c mice treated with PBS, or CAP. B Heatmaps showing that CAP upregulated genes involved in DNA damage (upper) and cytosolic DNA-sensing pathway (bottom). C qRT-PCR results showing that 5-FU upregulated the mRNA levels of Cgas, and Sting in 4T1 cells (upper) or 4T07 cells (bottom) stably expressing shNC or shTbkbp1. The data were represented as the mean ± SEM. Data were compared by using the Student’s t test. D qRT-PCR results showing that 5-FU upregulated the mRNA levels of Cgas, and Sting in AT3 cells (left) or TS/A cells (right) stably expressing Vector or Flag-Tbkbp1. The data were represented as the mean ± SEM. Data were compared by using the Student’s t test. E Western blot analysis showing that 5-FU increased the protein levels of Cgas, and Sting in 4T1 cells (left) or 4T07 cells (right) stably expressing shNC or shTbkbp1. Densitometric quantification of Western blots was performed using ImageJ software. F Western blot analysis showing that 5-FU increased the levels of Cgas, and Sting protein in AT3 cells (left) or TS/A cells (right) stably expressing Vector or Flag-Tbkbp1. Densitometric quantification of Western blots was performed using ImageJ software.

Article Snippet: The human embryonic kidney cell line HEK293T, human TNBC cell lines Hs 578 T, MDA-MB-468, BT-549, SUM159, LM2-4175, HCC1806, and the mouse TNBC cell line 4T1 were obtained from American Type Culture Collection (ATCC).

Techniques: RNA Sequencing, Quantitative RT-PCR, Stable Transfection, Expressing, Plasmid Preparation, Western Blot, Software

Journal: Oncogene

Article Title: TBKBP1 induces capecitabine resistance through negative regulation of type I interferon pathway in triple-negative breast cancer

doi: 10.1038/s41388-025-03598-4

Figure Lengend Snippet: A Viability of AT3 cells stably overexpressing Tbkbp1 (AT3-Tbkbp1, left) and TS/A cells stably overexpressing Tbkbp1 (TS/A-Tbkbp1, right) treated with DMSO (negative control), 5-FU (4 μ M), 5-FU (4 μM) with MG132 (10 μ M), 5-FU (4 μM) with BafA1 (200 nM), or 5-FU (4 μM) with 3-MA (5 mM) for 18 h, respectively. The data were represented as the mean ± SEM. Data were compared by using the Student’s t test. B Western blot analysis showing the expression of Tbk1 in AT3-Tbkbp1 cells (left) and TS/A-Tbkbp1 cells (right) treated with DMSO, 5-FU, or 5-FU plus the indicated inhibitors. Densitometric quantification of Western blots was performed using ImageJ software. C AT3-Tbkbp1 cells (left) and TS/A-Tbkbp1 cells (right) were treated with or without 200 nM BafA1 in combination of 4 μM 5-FU for the indicated times, and then subjected to Western blot analysis with Tbk1 and p62 antibodies. Densitometric quantification of Western blots was performed using ImageJ software. D Western blot analysis showing the expression of Tbk1, Irf3, and Ifngr1 when Tbkbp1-knockdown 4T1 cells (left) and 4T07 cells (right) were co-transfected with siTbk1 under 5-FU treatment for 18 h. Densitometric quantification of Western blots was performed using ImageJ software.

Article Snippet: The human embryonic kidney cell line HEK293T, human TNBC cell lines Hs 578 T, MDA-MB-468, BT-549, SUM159, LM2-4175, HCC1806, and the mouse TNBC cell line 4T1 were obtained from American Type Culture Collection (ATCC).

Techniques: Stable Transfection, Negative Control, Western Blot, Expressing, Software, Knockdown, Transfection