Hangeshashinto Induces Apoptosis and Suppresses Colorectal Cancer Growth In Vitro and In Vivo

Hangeshashinto (HST) is a standardized seven-herb Kampo pharmaceutical used to manage gastrointestinal symptoms and chemotherapy-associated mucosal toxicity, but its direct antitumor activity remains insufficiently characterized. We examined the effects of HST in DLD-1 and SW480 colorectal cancer (CRC) cells and in non-malignant fibroblast and colonic epithelial cell models. Cell viability, colony formation, apoptosis-related endpoints, gene expression, responses to 5-fluorouracil (5-FU), and the activity of representative constituents were evaluated. Oral antitumor efficacy and tolerability were assessed in female BALB/c-nu mice bearing DLD-1 xenografts. HST preferentially reduced CRC cell viability and colony formation and increased annexin V positivity, caspase-3 activity, cleaved caspase-3 staining, and apoptotic nuclear changes. z-VAD-FMK showed a nonsignificant tendency toward partial recovery of cell viability. Reverse transcription–quantitative polymerase chain reaction findings were exploratory; statistical significance was not assessed. Co-treatment with HST (0.3 mg/mL) and 5-FU significantly reduced cell viability compared with 5-FU alone in both DLD-1 and SW480 cells; pharmacological synergy was not evaluated. Oral HST inhibited xenograft growth; tolerability assessment was limited to body weight and serum biochemistry, with reduced serum uric acid in the high-dose group after washout. These findings provide a preclinical basis for further investigation of standardized HST in CRC.