Journal: Clinical & Experimental Metastasis

Article Title: A CSF-1R inhibitor both prevents and treats triple-negative breast cancer brain metastases in hematogenous preclinical models

doi: 10.1007/s10585-025-10366-x

Figure Lengend Snippet: CSF-1R suppression alters microglial cytokine secretion patterns with effects on astrocytes and tumor cells. A Mouse cytokine antibody array analysis of EOC2-derived conditioned medium treated with 100 nM BLZ945 or DMSO for 72 h. The images of cytokines with marked GM-CSF, IL-1Rα, IL-6 and TNFα are shown in the left panel. The pixel density of the above cytokines relative to the internal control is shown by the graphs in the right panel. B RT-PCR analysis of GM-CSF , IL-1Rα , IL-6 , and TNFα was performed on fixed, frozen specimens from vehicle control and BLZ945 treatment settings of the 4T1-BR5 TNBC brain metastasis model mice on Days -14, 3, and 10. Values represent the fold change ± SD relative to vehicle. Each dot represents one mouse, and the line designates the group median. C 4T1-BR5 and 231-BR cells were allowed to invade through Matrigel for 12 h in the presence of control or BLZ945 treated EOC2-derived conditioned medium. Invaded cells were visualized by Hema 3 staining and counted using ImageJ software. D and E EOC2 conditioned medium promotes the growth of 4T1-BR5 and 231-BR cells under co-culture conditions. 4T1-BR5 and 231-BR cells were labelled with mCherry and EGFP, respectively. Brain trophic cancer cell numbers are quantified and calculated by flow cytometry based on the ratio between cancer cells and quantitative beads. The effect of astrocytes alone on cancer cell growth was measured in low serum medium under co-culture conditions. n ≥ 3 biologically independent experiments. *, p < 0.05 vs DMSO control; n.s., not significant by the Mann–Whitney test. F Schematic representation of microglial action in breast cancer brain metastasis. CSF-1R + microglia/macrophages regulate the secretion of cytokines that influence the behavior of astrocytes and brain metastatic (BrM) cells within the microenvironment

Article Snippet: The mouse cytokine antibody array (#ARY006, R&D Systems) was used according to the manufacturer's instructions and analyzed using the ImageJ software.

Techniques: Ab Array, Derivative Assay, Control, Reverse Transcription Polymerase Chain Reaction, Staining, Software, Co-Culture Assay, Flow Cytometry, MANN-WHITNEY