Development of a compound herbal formulation (HBK) with antitumor and antioxidant functions for cancer adjuvant therapy

BACKGROUND: Cancer remains a leading cause of global mortality, with challenges such as metastasis, recurrence, and therapy resistance. Integrating traditional Chinese medicine with modern biomedicine has demonstrated the potential to enhance treatment efficacy. The tumor microenvironment (TME), particularly its immune landscape, plays a crucial role in cancer progression. PURPOSE: This study investigated HBK, a formulation of seventeen herbs and foods, for its potential to modulate the TME and boost antitumor immunity. METHODS: In a BALB/c lung cancer model, L1 tumor-bearing mice were fed either HBK-supplemented or control chow. Tumor growth, serum cytokines, and antioxidant enzymes were measured. Tumors and spleens were analyzed by flow cytometry, RNA sequencing, and Western blotting. RESULTS: HBK contains various bioactive compounds, including β-glucans, flavonoids, and polyphenols, which are known for their immunomodulatory properties. In a murine lung cancer model, HBK treatment significantly inhibited tumor growth and reduced lung metastasis. Serum levels of antioxidant enzymes (superoxide dismutase, glutathione peroxidase) and key antitumor cytokines (IL-12, IL-2, IFN-γ, MIP-1β) were significantly increased in HBK-treated mice. TME analysis revealed a shift toward an anti-tumor immune landscape, characterized by a reduction in pro-tumor immune cells (MDSCs, Th2, Th17, M2 macrophages, and Tregs) and an increase in cytotoxic natural killer (NK) and CD8⁺ T cells. RNA-Seq analysis identified upregulation of NK cell activation and chemokine signaling pathways, whereas Western blot analysis revealed downregulation of NF-κB and TLR3 pathways in the treated tumors. CONCLUSION: This study provides a comprehensive evaluation of a TCM-based multi-herb formulation for its immunomodulatory and antioxidant effects in lung cancer. HBK significantly enhanced anti-tumor immunity, increased antioxidant enzyme and cytokine activity, and remodeled the tumor microenvironment. Notably, these effects were mediated through inhibition of the TLR3/NF-κB signaling pathway. While individual herbal components have previously been linked to this pathway, our study is the first to demonstrate that a TCM-based multi-herb formula can modulate tumor immunity via TLR3/NF-κB inhibition in vivo.