murine colon carcinoma cell line ct26 cl25  (ATCC)

Journal: Frontiers in Immunology

Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model

doi: 10.3389/fimmu.2026.1733991

Figure Lengend Snippet: TAMpep-IP suppresses tumor growth in the colon cancer model. (A) BALB/c mice were subcutaneously inoculated with CT26 colon carcinoma cells (3 × 10 5 cells per mouse). Starting on day 7 post-inoculation, TAMpep-IP (400 nmol/kg) was administered subcutaneously every three days for a total of seven doses. (B) Representative images of tumors excised at the experimental endpoint (day 25) showed visibly reduced tumor size in the TAMpep-IP–treated group compared to control. (C) Tumor volumes were measured every 3 days following tumor implantation. Mice treated with TAMpep-IP exhibited significantly reduced tumor growth relative to the control group (control: n = 6; TAMpep-IP: n = 6). (D) Tumor proliferation was evaluated by immunohistochemical staining of Ki-67 in tumor sections. Quantitative analysis showed a significantly lower proportion of Ki-67 + proliferating cells in TAMpep-IP–treated tumors. Representative immunohistochemistry images were acquired at ×100 magnification. Scale bar = 1000 μm. All data are presented as mean ± SEM. *p<0.05, ***p<0.001.

Article Snippet: The murine colon carcinoma cell line CT26 (KCLB 80009; Korean Cell Line Bank, Seoul, Korea) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) (D5671; Welgene, Gyeongsangbuk, Korea) supplemented with 10% heat-inactivated FBS, 100 U/mL penicillin, and 1% penicillin–streptomycin.

Techniques: Control, Tumor Implantation, Immunohistochemical staining, Staining, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model

doi: 10.3389/fimmu.2026.1733991

Figure Lengend Snippet: TAMpep-IP reduces M2 macrophages in tumor tissues of colon cancer model. (A) Tumor-infiltrating immune cells were isolated from CT26 tumors in control and TAMpep-IP–treated mice. Flow cytometry was used to identify CD206 + F4/80 + macrophages within the CD45 + CD11b + population. TAMpep-IP significantly decreased the proportion of M2-like tumor-associated macrophages. (B) Quantitative RT-PCR analysis of tumor tissues revealed that TGF-β mRNA expression, a key M2-associated cytokine, was significantly reduced in TAMpep-IP–treated tumors compared to controls. (C) Western blot analysis of tumor showed a marked decrease in CD206 protein levels following TAMpep-IP, indicating effective suppression of M2 macrophage markers. (D) CD206 + macrophages were further visualized by immunohistochemical staining of tumor sections. ImageJ-based quantification confirmed a significant reduction in CD206 + area in TAMpep-IP–treated tumors. Representative immunohistochemistry images were acquired at ×100 magnification. Scale bar = 1000 μm. All data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The murine colon carcinoma cell line CT26 (KCLB 80009; Korean Cell Line Bank, Seoul, Korea) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) (D5671; Welgene, Gyeongsangbuk, Korea) supplemented with 10% heat-inactivated FBS, 100 U/mL penicillin, and 1% penicillin–streptomycin.

Techniques: Isolation, Control, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemical staining, Staining, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model

doi: 10.3389/fimmu.2026.1733991

Figure Lengend Snippet: TAMpep-IP enhances inflammatory cytokine expression and activated CD8 + T cells in tumor tissues of colon cancer model. (A) Flow cytometry was used to evaluate CD8 + T cell function in CT26 tumor tissues from control and TAMpep-IP-treated mice. TAMpep-IP significantly increased the proportion of activated Granzyme B + CD8 + T cells, while reducing the frequency of exhausted Tim-3 + CD8 + T cells, indicating enhanced cytotoxic T cell activity. (B, C) Confocal immunofluorescence analysis was performed on tumor sections stained with DAPI (nuclei), anti-CD8 (green), anti-Granzyme B (red), and anti-PD-1 (red). Activated CD8 + T cells were identified by co-localization of CD8 and Granzyme B, whereas exhausted CD8 + T cells were identified by co-localization of CD8 and PD-1. Quantification revealed a significant increase in intertumoral CD8 + Granzyme B + T cells and a concomitant decrease in CD8 + PD-1 + exhausted T cells following TAMpep-IP. Representative confocal images were acquired using a 40× objective lens. Scale bar = 20 μm. (D) Quantitative RT-PCR analysis of CT26 tumor tissues showed significantly elevated mRNA levels of pro-inflammatory cytokines TNF-α, IL-1β, and IL-12 in TAMpep-IP–treated mice compared to controls, indicating induction of a pro-inflammatory tumor microenvironment. All data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p < 0.0001.

Article Snippet: The murine colon carcinoma cell line CT26 (KCLB 80009; Korean Cell Line Bank, Seoul, Korea) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) (D5671; Welgene, Gyeongsangbuk, Korea) supplemented with 10% heat-inactivated FBS, 100 U/mL penicillin, and 1% penicillin–streptomycin.

Techniques: Expressing, Flow Cytometry, Cell Function Assay, Control, Activity Assay, Immunofluorescence, Staining, Quantitative RT-PCR

Journal: RSC Advances

Article Title: Exploring an azo-uracil based nickel( ii ) complex for anticancer and phosphatase like activities

doi: 10.1039/d6ra01587e

Figure Lengend Snippet: Cytotoxic and anti-cancer activity of the N1: (A) dose-dependent inhibition of cell proliferation in murine colon carcinoma cell lines (CT26 and MC-38) and human colorectal cancer cell line (HCT-15) upon treatment with increasing concentrations of N1, determined by MTT assay; (B) cell viability of normal murine fibroblast cells (NIH-3T3) following treatment with N1 at indicated concentrations, demonstrating minimal cytotoxicity; (C–E) dose–response curves showing percentage growth inhibition and corresponding IC 50 values for CT26 (C), MC-38 (D), and HCT-15 (E) cells. IC 50 values are calculated by nonlinear regression analysis using GraphPad Prism. Data are expressed as mean ± SD ( n = 3).

Article Snippet: Murine colon carcinoma cell lines CT26 and MC-38, human colorectal carcinoma cell line HCT-15, and normal murine fibroblast cell line NIH-3T3 were procured from the ATCC, USA.

Techniques: Activity Assay, Inhibition, MTT Assay

Journal: RSC Advances

Article Title: Exploring an azo-uracil based nickel( ii ) complex for anticancer and phosphatase like activities

doi: 10.1039/d6ra01587e

Figure Lengend Snippet: N1 induced apoptosis in colon cancer cells: (A) representative bright-field and fluorescence micrographs of CT26 cells treated with N1 (0.04 and 0.08 µg mL −1 ) for 12 h and stained with Annexin V-FITC (green) and propidium iodide (PI, red); merged images indicate apoptotic cell populations; (B–E) flow cytometric dot plots of Annexin V-FITC/PI staining showing viable (Q4, Annexin V − /PI − ), early apoptotic (Q3, Annexin V + /PI − ), late apoptotic (Q2, Annexin V + /PI + ), and necrotic (Q1, Annexin V − /PI + ) cell populations in untreated control (B), doxorubicin-treated positive control (0.25 µM) (C), and N1-treated cells at 0.04 µg mL −1 (D) and 0.08 µg mL −1 (E); (F) overlay histogram showing fluorescence intensity shifts in the FITC channel for control, doxorubicin, and N1-treated cells; (G) quantitative analysis of percentage dead/apoptotic cells under different treatment conditions (mean ± SD, n = 3; *** p < 0.001 vs. control).

Article Snippet: Murine colon carcinoma cell lines CT26 and MC-38, human colorectal carcinoma cell line HCT-15, and normal murine fibroblast cell line NIH-3T3 were procured from the ATCC, USA.

Techniques: Fluorescence, Staining, Control, Positive Control

Journal: RSC Advances

Article Title: Exploring an azo-uracil based nickel( ii ) complex for anticancer and phosphatase like activities

doi: 10.1039/d6ra01587e

Figure Lengend Snippet: Mechanistic insights into N1-induced apoptosis: caspase dependence, ROS generation, and mitochondrial perturbation: (A) representative flow cytometric dot plots of Annexin V-FITC/PI staining in CT26 cells treated with N1 (0.04 and 0.08 µg mL −1 ) for 12 h, showing viable (Annexin V − /PI − ), early apoptotic (Annexin V + /PI − ), late apoptotic (Annexin V + /PI + ), and necrotic (Annexin V − /PI + ) populations; (B) effect of pan-caspase inhibitor Z-VAD-FMK (20 µM, 1 h pre-treatment) on N1-induced apoptosis, showing reduced Annexin V-positive cell populations; (C) flow cytometric histograms of intracellular ROS levels measured using CellROX Green (total ROS) and MitoSOX Red (mitochondrial superoxide) in untreated, doxorubicin-treated (0.25 µM), and N1-treated cells (0.04 and 0.08 µg mL −1 ), indicating predominant induction of total ROS; (D) assessment of mitochondrial membrane potential (Δ Ψ m) using JC-1 staining, showing changes in red (JC-1 aggregates) and green (JC-1 monomers) fluorescence in control, N1-treated, and doxorubicin-treated cells, indicative of moderate mitochondrial depolarization. All experiments were performed in CT26 cells and analysed by flow cytometry. Data shown are representative of three independent experiments.

Article Snippet: Murine colon carcinoma cell lines CT26 and MC-38, human colorectal carcinoma cell line HCT-15, and normal murine fibroblast cell line NIH-3T3 were procured from the ATCC, USA.

Techniques: Staining, Membrane, Fluorescence, Control, Flow Cytometry

Journal: RSC Advances

Article Title: Exploring an azo-uracil based nickel( ii ) complex for anticancer and phosphatase like activities

doi: 10.1039/d6ra01587e

Figure Lengend Snippet: Effect of ROS scavenging and phosphatase inhibition on N1-induced cytotoxicity: (A) percentage cytotoxicity in CT26 cells treated with N1 (0.04 and 0.08 µg mL −1 ) in the presence or absence of N -acetyl cysteine (NAC, 5 mM) and PhosSTOP phosphatase inhibitor (1×); (B) corresponding cytotoxicity in HCT-15 cells under similar treatment conditions. NAC pre-treatment significantly reduces N1-induced cytotoxicity, indicating the involvement of ROS, while PhosSTOP treatment partially attenuates cytotoxicity, suggesting a possible contribution of phosphate-related processes. Data are expressed as mean ± SD ( n = 3; *** p < 0.001, **** p < 0.0001 vs. N1-treated group).

Article Snippet: Murine colon carcinoma cell lines CT26 and MC-38, human colorectal carcinoma cell line HCT-15, and normal murine fibroblast cell line NIH-3T3 were procured from the ATCC, USA.

Techniques: Inhibition