Exploring the Antitumor Mechanisms of Bai-Hua-She-She-Cao and Ban-Zhi-Lian against Head and Neck Squamous Cell Carcinoma: A Study on Natural Anticancer Therapeutics

Head and neck squamous cell carcinoma (HNSCC) is a highly lethal cancer with a poor prognosis. This research assessed the in vitro effectiveness and molecular mechanisms of Bai-Hua-She-She-Cao (BHSSC) and Ban-Zhi-Lian (BZL), a traditional Chinese medicinal herb pair against HNSCC. Bioactive compounds and targets of BHSSC-BZL were identified using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) and Swiss Target Prediction databases and cross-referenced with HNSCC-related targets from GeneCards and MalaCards. Network topology analysis identified hub genes, and The Cancer Genome Atlas (TCGA) data determined the protective and risk factors. Molecular docking verified target specificity, and in vitro experiments assessed BHSSC-BZL effects on CAL27 cell proliferation, apoptosis, and signaling pathways, with validation in SCC-25 cells. The study identified 21 bioactive compounds and 166 potential anti-HNSCC targets, highlighting 22 hub genes associated with cancer pathways, including phosphatidylinositol3-kinase/protein kinase b (PI3K/AKT) signaling and apoptosis regulation. Based on the TCGA data, ATP binding cassette transporter 1 (ABCB1) was identified as a protective factor, while the mesenchymal–epithelial transition factor (MET) was confirmed as a risk factor in HNSCC. Molecular docking analysis revealed that the MET protein could form stable complexes with seven candidate compounds derived from BHSSC-BZL. In vitro experiments further demonstrated that BHSSC-BZL significantly inhibited CAL27 and SCC-25 cell proliferation. BHSSC-BZL demonstrates anti-HNSCC potential by downregulating MET and Bcl-2, upregulating Bax and cleaved-Caspase-9, and inhibiting the PI3K/AKT/mTOR signaling. BHSSC-BZL demonstrates anti-HNSCC activity via a multicomponent, multitarget, and multipathway mechanism, highlighting its potential as a natural antitumor agent.