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: ACS Omega

Article Title: Feasibility of a Protease Activity-Based Nanosensor for Breast Cancer Screening

doi: 10.1021/acsomega.5c09454

Figure Lengend Snippet: Catalytic efficiency of MMP-2 on fluorogenic probes and assessment of the ABN toxicity. (a) Schematic showing the catalytic efficiency of MMP-2 on TIAH–LH, immobilized on the surface of microplate wells and incubated with the 50 nM catalytic domain for 2 h at 37 °C. Initial cleavage velocity of the substrate conjugated onto the nanocarrier surface at (b) 32.5 Å and (c) 53.4 Å from the core, highlighting the catalytic efficiency. The lines represent the Michaelis–Menten model fit to the data. mean ± s.e.m. is denoted by a horizontal line with error bars; n = 2 wells per probe concentration; N = 3; R 2 = 0.87; R 2 = 0.99. K cat , catalytic constant; K M , Michaelis–Menten constant. (d–g) Blood levels of hepatotoxicity markers of the mice injected with the ABN were equal to the levels found in the controls. The mean body weight remained stable for 5 d after ABN injection. Each square represents one mouse, and the mean value is denoted by the red bar. The Shapiro–Wilk test is followed by the unpaired two-tailed t -test; * P > 0.05. ALT, alanine aminotransferase; AST, aspartate aminotransferase. (h, i) Viability of 4T1 and ID8 cells after incubation with the ABN at different concentrations for 24 h. mean ± s.e.m. is denoted by a horizontal line with error bars; n = 2–3 wells per condition; N = 3; one-way ANOVA with Bonferroni multiple comparisons; ** P < 0.05. Ctrl, positive control: acetaminophen at 25 mM in cell culture media.

Article Snippet: Mouse breast cancer cell 4T1 was cultured in ATCC-formulated RPMI medium supplemented with 10% fetal bovine serum and mouse ovarian surface epithelial cell ID8 in high glucose DMEM supplemented with 4% fetal bovine serum, 5 μg mL –1 insulin and transferrin, and 5 ng mL –1 sodium selenite.

Techniques: Incubation, Concentration Assay, Injection, Two Tailed Test, Positive Control, Cell Culture

Journal: ACS Omega

Article Title: Feasibility of a Protease Activity-Based Nanosensor for Breast Cancer Screening

doi: 10.1021/acsomega.5c09454

Figure Lengend Snippet: ABN performance in the 4T1 mammary tumor model. (a) Kinetics of the ABN signal in the plasma of mice bearing a four-week-old tumor. The lines denote the behavior predicted by a one-phase exponential decay model. Values are represented as mean ± s.e.m; n = 2–4 mice; R 2 = 0.99, tumor; R 2 = 0.98, nontumor. (b) Kinetics of the biomarker signal in the plasma of tumor-bearing mice versus normal counterparts, indicating the exacerbation of protease activity in the blood. Lines denote the kinetic behavior by a one-phase exponential decay model. Values are represented as mean ± s.e.m.; n = 2–4 mice; R 2 = 0.78, tumor; R 2 = 0.74, nontumor.

Article Snippet: Mouse breast cancer cell 4T1 was cultured in ATCC-formulated RPMI medium supplemented with 10% fetal bovine serum and mouse ovarian surface epithelial cell ID8 in high glucose DMEM supplemented with 4% fetal bovine serum, 5 μg mL –1 insulin and transferrin, and 5 ng mL –1 sodium selenite.

Techniques: Clinical Proteomics, Biomarker Discovery, Activity Assay

Journal: Exploration

Article Title: A Groundbreaking Electric Field‐Induced Cascade Gas Therapy Against Large Volume Solid Tumor Through Electro‐Stress Storm

doi: 10.1002/EXP.20240410

Figure Lengend Snippet: In vitro EGT performance and cytotoxicity of PR@L. (A) CLSM observation of RhB‐labeled PR internalized by 4T1 cells after different culture time. scale bar: 10 µm. (B) Schematic diagram of the experimental device for cytotoxicity evaluation of EGT system. (C) Cytotoxicity of 4T1 cells cultured with various formulations at diverse concentrations for 24 h. (D) Relative cytotoxicity of EGT‐treated 4T1 cells after incubation with different formulations (at 100 µg mL −1 Pt concentration) and (E) PR@L under various square‐wave AC parameters (10 mHz). (F) Intracellular RNS production after diverse treatments with or without electrical stimulation monitored by CLSM (scale bar: 50 µm) and (G) flow cytometer. (H) Intracellular ROS (DCF) and NO (DAF‐FM‐NO) CLSM images and corresponding fluorescence intensity profiles along the arrow (scale bar: 50 µm). n = 3 and * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: 4T1 mouse breast cancer cell line was provided by the American Type Culture Collection.

Techniques: In Vitro, Labeling, Cell Culture, Incubation, Concentration Assay, Flow Cytometry, Fluorescence

Journal: Exploration

Article Title: A Groundbreaking Electric Field‐Induced Cascade Gas Therapy Against Large Volume Solid Tumor Through Electro‐Stress Storm

doi: 10.1002/EXP.20240410

Figure Lengend Snippet: In vivo EGT treatment for solid tumor with large size by electro‐stimulating gas donor‐encapsulated PR. (A) Schematic illustration of in vivo EGT treatment operation. (B) Tumor growth curves of 4T1 tumor‐bearing mice after different treatments ( n = 5). (C) Average tumor weights and (D) mouse weight at the end of the experiment (at day 10, n = 5). (E) Tumor growth curves of individual mouse from each group. (F) Ki67, H&E, and TUNEL staining analyses of tumor tissue slices collected from different groups after treatment. Scale bar: 50 µm. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: 4T1 mouse breast cancer cell line was provided by the American Type Culture Collection.

Techniques: In Vivo, TUNEL Assay, Staining

Journal: Exploration

Article Title: A Groundbreaking Electric Field‐Induced Cascade Gas Therapy Against Large Volume Solid Tumor Through Electro‐Stress Storm

doi: 10.1002/EXP.20240410

Figure Lengend Snippet: EGT‐induced in vivo immunogenic effect boosted tumor immunotherapy. (A) Scheme illustration of immune activation mechanisms of the EGT platform through releasing immunogenic DAMPs and generating PR@L‐mediated electro‐stress storm. (B) Immunofluorescence analysis for in situ CRT exposure in 4T1 tumors from mice after various treatments. Scale bar: 100 µm. (C) Flow cytometric analysis and (D) corresponding quantitative analysis ( n = 3) of DCs maturation (gated on CD11c + CD80 + CD86 + ) in tumor draining lymph node cells collected from mice 10 days after treatments. (E) Immunofluorescence analysis (scale bar: 100 µm), (F) flow cytometric analysis, and (G) corresponding quantitative analysis ( n = 3) of infiltrating CD8 + T cells (gated on CD45 + CD3 + CD8 + ) in tumor tissues from mice after various treatments. (H) ELISA analysis of cytokines in serum from 4T1 tumor‐bearing mice, including TNF‐α, INF‐γ, and IL‐10 ( n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: 4T1 mouse breast cancer cell line was provided by the American Type Culture Collection.

Techniques: In Vivo, Activation Assay, Immunofluorescence, In Situ, Enzyme-linked Immunosorbent Assay