Scorpio fuscus venom as a promising anticancer agent against colorectal cancer.

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, underscoring the urgent need for novel and effective therapeutic agents. Conventional treatments, including chemotherapy, radiotherapy, and surgery, are often associated with significant adverse effects, prompting the exploration of alternative therapeutic strategies. This study aimed to evaluate the anticancer effects of Scorpio fuscus venom (SFV) on human colorectal cancer models using integrated in vitro and in vivo approaches. SFV constituents were characterized using gel electrophoresis, followed by high-performance liquid chromatography and UV-visible spectrometry. Identified peptides were subjected to structural modeling and in silico docking analyses against selected proteins associated with colorectal cancer and apoptosis-related pathways. The cytotoxic effects of SFV were assessed in human CRC cell lines (DLD-1, HT-29, and CaCo-2) and a healthy colon epithelial cell line (CCD-18Co) using Alamar Blue assays after 48-h treatment, and half-maximal inhibitory concentration (IC₅₀) values were determined. SFV treatment resulted in a dose-dependent reduction in cancer cell viability, accompanied by decreased migratory capacity and colony formation ability. Apoptotic responses were further evaluated by flow cytometry and gene expression analyses, indicating modulation of apoptosis-associated genes. For in vivo validation, subcutaneous and orthotopic xenograft colon cancer models were established in mice. SFV administration led to reduced tumor growth compared with control groups. Immunohistochemical analyses revealed altered expression patterns of selected tumor-related markers in SFV-treated tumors. Gene expression profiling demonstrated ≥ twofold changes in 51 genes, including downregulation of BAK1 and TRAF3, and upregulation of BIRC2, BIRC3, BIRC6, CASP8, TNFRSF8, TNFRSF11, and BOK. Collectively, these findings indicate that SFV exerts significant antitumor effects in colorectal cancer models and support its potential as a promising anticancer agent, warranting further mechanistic and translational investigation.