Journal: Frontiers in Immunology
Article Title: A novel hollow iron nanoparticle system loading PEG-Fe 3 O 4 with C5a receptor antagonist for breast cancer treatment
doi: 10.3389/fimmu.2024.1466180
Figure Lengend Snippet: Evaluations of anti-tumor effect of mesoporous hollow PEG-Fe 3 O 4 . (A-D) , Cell viability of mouse breast cancer cell line 4T-1, non-malignant mouse fibroblast cell line HC11, human breast cancer cell line MCF-7, human normal mammary epithelial cell line MCF-10A were determined after treatment with PEG-Fe 3 O 4 for 48h (n=4). Statistical differences are assessed by one-way ANOVA. Data were mean ± s.e.m. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.001. (E) , ROS level analysis of 4T-1 cells treated with PEG-Fe 3 O 4 (200ug/mL) for 48h (n=3). (F) , Immunoblotting of NRF2 and GPX4 in 4T-1 cells after treatment with PEG-Fe 3 O 4 for 48h. (G-I) , mice were implanted with 1.0 × 10 6 4T-1 cells in the mammary fat pad with and without PEG-Fe 3 O 4 treatment. The tumor growth was measured by representative tumor images (G) , tumor weight (H) , and tumor size (I) (n=6). (J-L) , Immunohistochemistry and and statistical analyses of the GPX4 and NRF2 expression in tumor tissues from 4T-1 xenografts and 4T-1 xenografts with PEG-Fe 3 O 4 treatment. (M) , Quantification of complement activation products C5a by ELISA: levels of anaphylatoxin C5a in mouse plasma (administered with PBS or PEG-Fe 3 O 4 ) (n=10). (N, O) , Immunohistochemistry and statistical analyse of C5b-9 in tumor tissues from 4T-1 xenografts (left) and 4T-1 xenografts (right) with PEG-Fe 3 O 4 . (P-S) , Expressions of M1 associated gene (iNOS, CCR7) and M2 associated gene (Arg1, CCL17) were measured by RT-qPCR in RAW264.7 and THP-1 treated with PEG-Fe 3 O 4 for 48h, and the expression levels of mRNA were normalized to β-actin (n=3). (T, U) , Flow cytometry analysis of IA-IE and CD206 expression in tumor-associated macrophages, representative flow cytometry diagram (T) and the ratio of M1/M2 macrophages (U) . Statistical differences are assessed by student’s t-test. Data were mean ± s.e.m. *, p < 0.05; **, p < 0.01; ***, p < 0.001.
Article Snippet: The mouse 4T-1 and HC11 cell lines were obtained from Procell Life Science and Technology.
Techniques: Western Blot, Immunohistochemistry, Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Quantitative RT-PCR, Flow Cytometry
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