Integrative Computational and Experimental Analysis Reveals Piperlongumine Induces Reactive Oxygen Species-Mediated Apoptosis and Targets Hub Genes in Colorectal Cancer

Abstract

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. Multiple therapeutic strategies including chemotherapy are being followed in CRC therapy. But, prolonged dependency on chemotherapy leads to drug resistance. Growing evidence suggests that phytocompounds could offer safer and effective therapeutic options. This study investigates the anticancer potential of Piperlongumine (PIP), a natural phytocompound, against CRC using an integrated bioinformatics and experimental approach. By the initial integration and analysis of multiple databases, 11 common differentially expressed genes were identified that may serve as potential targets of PIP in CRC, many of which are involved in key signalling and apoptotic pathways. Protein-protein interaction analysis revealed five major hub genes- TP53, CCND1, AKT1, CTNNB1, and IL1B that were significantly linked to poor prognosis and metastasis in CRC. Molecular docking revealed a strong binding of PIP to these targets, along with favorable pharmacokinetic properties. In-vitro validations using CRC cell lines (SW-480 and HT-29) revealed dose-dependent cytotoxicity with IC50 values of 3 μM and 4 μM, respectively. Further, PIP inhibits migration, induces DNA damage, and promotes reactive oxygen species (ROS)-mediated apoptosis as well as cell cycle arrest. In addition, PIP modulates hub gene expression (TP53 ↑; CCND1, AKT1, CTNNB1, and IL1B ↓), supporting its mechanistic role in CRC. Overall, our findings suggest that PIP exerts strong anticancer activity in CRC by inducing ROS-mediated apoptosis and targeting critical genes involved in cancer progression, suggesting its potent role as a promising anticancer agent.