Chelidonine-induced inhibition of FBP1 disrupts M2 macrophage polarization and attenuates breast cancer

Background: Chelidonium majus l., a traditional herbal medicine listed in the Pharmacopoeia of China (2025), has remarkable anticancer properties. Chelidonine (CHE), its primary bioactive alkaloid, can inhibit the proliferation of breast cancer (BC) cells; however, whether its antitumor effect in BC is mediated through regulating M2 macrophage polarization remains unexplored.
Purpose: To investigate the effect of CHE on M2 macrophage polarization in BC and to identify key molecular targets and pathways involved in its immunomodulatory effects.
Methods: The effect of CHE on tumour progression was evaluated in vivo using a 4T1 BCE mouse model. In addition, the effects of CHE on M2 macrophage polarization were investigated both in vitro and in vivo via immunofluorescence and Western blotting. Coculture assays were conducted to elucidate the mechanism by which CHE exerts antiproliferative and antimetastatic effects through the inhibition of M2 macrophage polarization. An integrated analysis of network pharmacology, single-cell, and bulk RNA-seq data was performed to identify potential molecular targets of CHE in the suppression of M2 macrophage polarization. Molecular docking, Western blotting, and ELISA were utilized for target validation to elucidate the roles in M2 macrophage
polarization.
Results: CHE effectively impeded tumour progression in BC mice. Both in vitro and in vivo experiments demonstrated that CHE significantly inhibited M2 macrophage polarization, as evidenced by the altered expression of key polarization markers, including CD86, CD206, Arg-1, and CD11c. Under coculture conditions, CHE disrupted the mitochondrial membrane potential and compromised membrane integrity in BC cells, triggering pyroptosis via activation of the NLRP3/Caspase-1/GSDMD signaling axis. Additionally, CHE attenuated the epithelial–mesenchymal transition (EMT) process, thus reducing the migratory and invasive capacities of BC cells. An integrated analysis of network pharmacology, single-cell, and bulk RNA sequencing data revealed FBP1 as a potential molecular target through which CHE suppresses M2 macrophage polarization. Molecular docking studies revealed a relatively strong binding affinity between CHE and FBP1. Experimental validation further confirmed that CHE downregulated the expression of FBP1 and its downstream target p-STAT3 in macrophages, thereby inhibiting M2 polarization.
Conclusions: CHE might suppress M2 polarization and tumour progression by targeting FBP1. These findings highlight its potential as a therapeutic agent in immunotherapy and BC treatment.