Identification of molecular therapeutic features of the homeopathy medicine Thuja by genome-wide expression profiling

• Thuja occidentalis reduces proliferation, colony- and spheroid-forming features of gastric cancer cells. • Thuja downregulates multiple oncogenic pathways including MAPK, MYC, WNT, NOTCH, GPCR, TGF-β, PDGF, and JAK/STAT. • In concordance with Thuja ’s traditional use in treating warts, T. occidentalis downregulates the genes involved in the pathogenesis of warts. • Thujone, a bioactive compound of T. occidentalis , also exhibits cytotoxic and therapeutic features. Thuja occidentalis , a homeopathic remedy, has been extensively used in traditional medicine for treating various ailments, particularly for the skin disease warts. Its therapeutic use is well-documented, especially in Chinese traditional medicine, where it has also been explored for its potential in cancer treatment. In Traditional Chinese Medicine (TCM), Thuja occidentalis is known as “ Ce Bai Ye” (崖柏屬 ) and has been extensively employed due to its diverse therapeutic features. Ce Bai Ye is particularly valued for its hemostatic, astringent, and anti-inflammatory properties. In traditional Chinese medicine Thuja's astringent nature makes it beneficial for digestive issues such as diarrhoea. Despite the above-mentioned potential applications, the underlying molecular mechanisms remain to be identified. This study was designed to investigate the molecular impact of Thuja on cancer cells, with a special focus on its effects on gastric cancer cells. This study aims to investigate the molecular therapeutic features of Thuja occidentalis in gastric cancer by employing genome-wide expression profiling and by evaluating the impact on the growth and signalling pathways in gastric cancer cells. AGS, gastric cancer cells were treated with 0.1% Thuja occidentalis mother tincture, and the impact on inhibiting the features of cancer cell growth was assessed by colony and spheroid-forming assays. Genome-wide expression profiling was conducted to identify the genes and pathways regulated by Thuja . Gene set enrichment analysis was done to elucidate the signalling pathways modulated by Thuja . Thuja has significantly inhibited the growth of gastric cancer cells and also reduced the colony- and spheroid-forming potential. Genome-wide expression profiling has identified a significant downregulation of histone and zinc finger (ZNF) family genes, polymerase-related genes, and ATP genes. Gene set enrichment analysis revealed Thuja to downregulate the signalling pathways, including MAPK, MYC, Wnt, NOTCH, GPCR, TGF-β, PDGF, and JAK/STAT. The genes commonly upregulated in warts were downregulated by Thuja , indicating a potential therapeutic role of Thuja in wart treatment. This study provides novel insights into the molecular therapeutic effects of Thuja in gastric cancer cells. The data also provides the lead knowledge for the further development of Thuja as a targeted therapeutic agent for gastric cancer and warrants further pre-clinical investigations.