Aiduqing formula inhibits breast cancer metastasis by suppressing TAM/CXCL1-induced Treg differentiation and infiltration

Abstract

Background Metastasis represents the leading cause of death in patients with breast cancer. Traditional Chinese medicine is particularly appreciated for metastatic diseases in Asian countries due to its benefits for survival period prolongation and immune balance modulation. However, the underlying molecular mechanisms remain largely unknown. This study aimed to explore the antimetastatic effect and immunomodulatory function of a clinical formula Aiduqing (ADQ). Methods Naive CD4+ T cells, regulatory T cells (Tregs), and CD8+ T cells were sorted by flow cytometry. Then, breast cancer cells and these immune cells were co-cultured in vitro or co-injected into mice in vivo to simulate their coexistence. Flow cytometry, ELISA, qPCR, double luciferase reporter gene assay, and chromatin immunoprecipitation assay were conducted to investigate the immunomodulatory and antimetastatic mechanisms of ADQ. Results ADQ treatment by oral gavage significantly suppressed 4T1-Luc xenograft growth and lung metastasis in the orthotopic breast cancer mouse model, without noticeable hepatotoxicity, nephrotoxicity, or hematotoxicity. Meanwhile, ADQ remodeled the immunosuppressive tumor microenvironment (TME) by increasing the infiltration of tumor-infiltrating lymphocytes (TILs) and cytotoxic CD8+ T cells, and decreasing the infiltration of Tregs, naive CD4+ T cells, and tumor-associated macrophages (TAMs). Molecular mechanism studies revealed that ADQ remarkably inhibited CXCL1 expression and secretion from TAMs and thus suppressed the chemotaxis and differentiation of naive CD4+ T cells into Tregs, leading to the enhanced cytotoxic effects of CD8+ T cells. Mechanistically, TAM-derived CXCL1 promoted the differentiation of naive CD4+ T cells into Tregs by transcriptionally activating the NF-κB/FOXP3 signaling. Lastly, mouse 4T1-Luc xenograft experiments validated that ADQ formula inhibited breast cancer immune escape and lung metastasis by suppressing the TAM/CXCL1/Treg pathway. Conclusions This study not only provides preclinical evidence supporting the application of ADQ in inhibiting breast cancer metastasis but also sheds novel insights into TAM/CXCL1/NF-κB/FOXP3 signaling as a promising therapeutic target for Treg modulation and breast cancer immunotherapy.

Integration analysis identifies MYBL1 as a novel immunotherapy biomarker affecting the immune microenvironment in clear cell renal cell carcinoma: Evidence based on machine learning and experiments

“…In ccRCC, Ji et al found that the hamper of Tregs in the colon cancer microenvironment can improve the anti-tumor effect and inhibit cancer metastasis (29). Li et al indicated that aiduqing formula can inhibit Treg infiltration induced by TAM/CXCL1, further hampering breast cancer metastasis (30). In solid tumors, M2 macrophages are generally cancer promoters (31).…”

“…Li et al. indicated that aiduqing formula can inhibit Treg infiltration induced by TAM/CXCL1, further hampering breast cancer metastasis ( 30 ). In solid tumors, M2 macrophages are generally cancer promoters ( 31 ).…”

Upregulation of Ferroptosis-Related Fanconi Anemia Group D2 is a Poor Prognostic Factor and Indicator of Tumor Immune Cell Infiltration in Lung Adenocarcinoma

“…Notably, LUAD tends to metastasize at an early stage, which results in a high mortality rate. Further, immune escape is the rst crucial step for tumor initiation, progression, and metastasis in TME [32]. Tumor-in ltrating immune cells (TIICs) are an integral part of the TME, playing important roles in suppressing or promoting tumor cell growth.…”

“…Tumor-in ltrating immune cells (TIICs) are an integral part of the TME, playing important roles in suppressing or promoting tumor cell growth. Therefore, remodeling the TME from immunesuppressive to immune-promoting could be a potential anti-tumor therapeutic for LUAD [32]. Recently, studies have shed light on the e cacy of immunotherapy.…”

Correction to: Aiduqing formula inhibits breast cancer metastasis by suppressing TAM/CXCL1-induced Treg differentiation and infiltration

“…Following publication of the original article [ 1 ], the authors found an error in Fig. 6 D. Unintentionally, the same lung was mistakenly displayed twice in the TAM group (the upper and the lower image).…”