normal mouse l929 fibroblast cell line  (ATCC)

Journal: Biomaterials

Article Title: Dual inhibition of oxidative phosphorylation and glycolysis using a hyaluronic acid nanoparticle NOX inhibitor enhanced response to radiotherapy in colorectal cancer.

doi: 10.1016/j.biomaterials.2025.123437

Figure Lengend Snippet: Fig. 1. Effects of HANP/GKT831 on cell proliferation and ROS production in tumor cell lines. A. Production of HANP/GKT831. B. SRB Cell Proliferation assay. IC50 of HANP/GKT831 in the MC38 tumor cell line was 0.007 μM (upper left), HT29 cell line was 5.505 μM (upper right), CCL227 cell line was 1.668 μM (lower left). Neither HANP/GKT831 nor GKT831 inhibited the proliferation of normal mouse fibroblast cells (NIH 3T3 cell line). C. The level of intracellular H2O2 production in the MC38 tumor and NIH 3T3 fibroblast cells. Intracellular ROS, H2O2, produced by NOX1 indirectly and NOX4 directly were detected by a cell-based DCFDA assay that was performed 60 h following treatment. D and E. Intracellular mitochondrial ROS determined by the measurement of H2O2 level (D: Mito PY1 assay) or O2 •−(E: Mito SOX assay) 60 h following treatment. Cells were treated with 0.01 μM of equivalent doses of GKT831 or HANP/GKT831. Fluorescence intensity (FI) represents the level of ROS in the treated cells. The mean FI of five samples is shown. ns, not significant. Student’s t-test: *p < 0.05, **p < 0.01, and ns: not significant, p > 0.05.

Article Snippet: Human CRC HT-29 and CCL227 cell lines, and normal mouse fibroblast cell line, NIH 3T3, were obtained from ATCC (Manassas, VA, USA).

Techniques: Sulforhodamine B Assay, Produced, Fluorescence

Journal: Biomaterials

Article Title: Dual inhibition of oxidative phosphorylation and glycolysis using a hyaluronic acid nanoparticle NOX inhibitor enhanced response to radiotherapy in colorectal cancer.

doi: 10.1016/j.biomaterials.2025.123437

Figure Lengend Snippet: Fig. 4. Targeted delivery and therapeutic response in the mouse CRC model. A. Targeted delivery of HANP/GKT831 in tumors following systemic adminis tration. Whole body and ex vivo NIR optical imaging detected targeted delivery and distribution at 24 h after i.v. injection. The mean fluorescent signals were analyzed and quantified from three mouse images. B. Immunofluorescence labeling of CD31+ tumor vessels (red arrows) and NIR 830-HANPs (green arrows). NIR 830-HANPs (green arrows) were colocalized with CD44+ tumor cells (red arrows). Immunofluorescence labeling of FAP + fibroblasts (red). Green arrows: NIR 830- HANPs. Scale bar = 50 μm. HANP/GKT831 treatment inhibited tumor growth and enhanced therapeutic response to RT in the mouse CRC model. C. Treatment protocol. The MC38 tumor bearing mice received i.v. injections of 5 mg/kg of GKT831 equivalent dose of HANP/GKT831. 2 Gy of RT was applied 24 h post the injection. D. Tumor growth curve. The mean tumor volumes of each group at different time points after treatment are shown. E. The average tumor weights of each group were collected 5 days after the last treatment. F. Monitoring body weights during treatment. The mean body weight of each mouse group is shown. n = 5 mice/ group. Student’s t-test: *p < 0.05, **p < 0.01 and ****p < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Human CRC HT-29 and CCL227 cell lines, and normal mouse fibroblast cell line, NIH 3T3, were obtained from ATCC (Manassas, VA, USA).

Techniques: Clinical Proteomics, Ex Vivo, Optical Imaging, Injection, Immunofluorescence, Labeling

Journal: Biomaterials

Article Title: Dual inhibition of oxidative phosphorylation and glycolysis using a hyaluronic acid nanoparticle NOX inhibitor enhanced response to radiotherapy in colorectal cancer.

doi: 10.1016/j.biomaterials.2025.123437

Figure Lengend Snippet: Scheme 1. Mechanism of HANP/GKT831 treatment in inhibition of ROS and metabolism signal pathways and enhancement of therapeutic responses to RT in cancer cells. Systemic delivery of HANP/GKT831 leads to the accumulation of GKT831 in the tumor cells through active CD44 targeting and HA homing to inflammatory tumor areas. The binding of HANP/GKT831 to CD44 receptors that are highly expressed in tumor cells results in the internalization into endosomes/ lysosomes for intracellular release of GKT831 mediated by hyaluronidase 1. A high concentration of GKT831 effectively inhibits the activity of NOX1 on the plasma membrane and endosomes and NOX4 on the mitochondrial, nuclear, and endoplasmic reticulum membrane, leading to significant decreases in ROS production, including superoxide anion (O2•-) and hydrogen peroxide (H2O2), causing redox stress in tumor cells. Reduced ROS suppresses both glycolysis and mitochondrial OXPHOS to decrease ATP production, induces bioenergetic stress, and upregulates inflammatory cytokines, contributing to an enhanced immune response. Concurrently, redox and bioenergetic stress downregulate signaling pathways involved in DNA repair, cell proliferation, and survival, thereby sensitizing tumor cells to RT. Additionally, the depletion of fibroblasts, M2-type TAMs, and immunosuppressive cytokines reprograms the TME, further promoting immune-mediated tumor cell death [36]. In summary, HANP/GKT831 not only disrupts tumor redox and metabolic homeostasis but also enhances the sensitivity of tumor cells to radiotherapy and promotes antitumor immunity.

Article Snippet: Human CRC HT-29 and CCL227 cell lines, and normal mouse fibroblast cell line, NIH 3T3, were obtained from ATCC (Manassas, VA, USA).

Techniques: Inhibition, Binding Assay, Concentration Assay, Activity Assay, Clinical Proteomics, Membrane, Protein-Protein interactions

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Royal jelly induces ROS-mediated apoptosis in acute lymphoblastic leukemia (ALL)-derived Nalm-6 cells: Shedding light on novel therapeutic approaches for ALL

doi: 10.22038/IJBMS.2024.76261.16498

Figure Lengend Snippet: Effect of royal jelly (RJ) on normal cells. L929 and PBMC were used as models of normal cells. The MTT assay indicated that RJ has no cytotoxic effect on L929 and PBMC cells (A). Analysis of apoptosis using Annexin-V/PI staining and flow cytometry revealed that RJ has no significant apoptotic effect on normal cells (B). Hemolytic activity evolution suggested that the hemolysis rate decreases when RJ concentration increases (C). The values are given as the mean SD of three independent experiments. * P ≤0.05 represents significant statistical changes from the control sample

Article Snippet: Cell culture and treatment Acute Lymphoblastic Leukemia cell line (Nalm-6) and normal mouse fibroblast cell line (L929) were purchased from the Pasteur Institute collection (Tehran, Iran).

Techniques: MTT Assay, Staining, Flow Cytometry, Activity Assay, Concentration Assay, Control