Mechanistic Investigation of Nitidine Chloride-Mediated Anti-Colorectal Cancer Activity: Centromere-Associated...

Colorectal cancer (CRC) is counted among the most widespread malignancies worldwide, characterised by elevated incidence and mortality rates. Conventional chemotherapy is frequently associated with severe toxic side effects and the development of drug resistance, which necessitates an urgent search for alternative therapeutic modalities. Traditional Chinese medicine (TCM), distinguished by its multi-component and multi-target synergistic actions, represents a promising prospect for the development of innovative anti-tumour therapies. Nitidine chloride (NC), a major bioactive component isolated from Zanthoxylum nitidum, has demonstrated notable anti-tumour activity in various cancer types. However, the specific molecular mechanisms underlying its anti-CRC effects remain unclear. Centromere-associated protein E (CENPE) exerts a pivotal function in the regulation of the cell cycle, and its aberrant expression has been documented in multiple malignancies. It may therefore serve as a potential therapeutic target. This study sought to clarify the interplay between NC and CENPE, with the aim of offering a scientific foundation for the advancement of precision therapeutic approaches for CRC utilising TCM-derived bioactive compounds. To comprehensively characterise the expression pattern of CENPE in CRC, we integrated a range of state-of-the-art technologies encompassing single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, large-scale mRNA cohort analyses and immunohistochemistry (IHC). The regulatory impact of NC on CENPE expression was verified through real-time quantitative polymerase chain reaction (RT-qPCR) and IHC. Additionally, molecular dynamics simulation (MDS) was employed to investigate the binding mode and stability of the NC-CENPE complex. Multi-dimensional analyses indicated that CENPE is significantly overexpressed in CRC tissues, with a standardised mean difference of 1.32, and its expression scores approach 1.0 in malignant regions. CRISPR screening data suggested that CENPE knockout is associated with markedly reduced proliferation of CRC cells. MDS data supported a plausible binding mode between NC and CENPE, with a predicted binding free energy of -8.2 kcal/mol, in which van der Waals interactions constituted a major component of the calculated binding energy. Furthermore, treatment with NC was associated with significant downregulation of CENPE mRNA and protein levels in CRC cells and xenograft models in this study, although these findings require further validation in additional experimental systems. NC exerts anti-colorectal cancer activity through targeting CENPE. This discovery lays a mechanistic foundation for the development of precision therapies based on active TCM ingredients, offering a new direction for CRC treatment.