ABSTRACT: Colorectal cancer (CRC) is one of the most prevalent and lethal malignancies worldwide. The epithelial-mesenchymal transition (EMT) plays a critical role in tumor invasion and metastasis. Piperine, a natural alkaloid, has demonstrated antitumor activity; however, its low bioavailability limits clinical application. HJJ_3_5, a novel piperine derivative synthesized and validated by our research group, exhibits potent antitumor effects against colorectal cancer cells. In addition to its strong cytotoxicity, HJJ_3_5 disrupts key metastatic processes, including DNA replication, cell migration, invasion, and adhesion, showcasing its multifaceted mechanism of action. In vivo, HJJ_3_5 shows significant efficacy in the CAM model, effectively inhibiting tumor growth and angiogenesis, underscoring its potential as a powerful anti-cancer agent. At the molecular level, comprehensive transcriptomic analysis reveals that HJJ_3_5 significantly downregulates the EMT pathway. Notably, machine learning algorithms identify six core EMT-related genes (COL12A1, PJA2, VCAN, MEF2C, DPYD, and DDR2) as pivotal drivers of cancer progression, with their expression strongly associated with poor clinical outcomes. Single-cell RNA sequencing further reveals that these genes are predominantly expressed in fibroblasts and myofibroblasts, key players in the EMT process and metastasis. These findings position HJJ_3_5 as a promising therapeutic candidate, offering a targeted and potent approach to halting the relentless progression of colorectal cancer.
