Iphigenia indica Kunth extract induces apoptosis in colorectal cancer cells via PIK3CD and AKT/mTOR pathway

This study explored the anti-cancer mechanisms of Iphigenia indica Kunth extract in human colorectal cancer (CRC) SW480 cells, focusing on its potential to induce apoptosis by inhibiting PIK3CD and modulating the AKT/mTOR transduction machinery. Network pharmacology facilitated the identification of active molecules, specifically ellagic acid, stigmasterol, beta-sitosterol, and 2-methoxy-9,10-dihydrophenanthrene-4,5-diol, and their potential targets. In vitro assays with SW480 cells were carried out to screen the extract's effects on cell growth, migration, and apoptotic death. Key targets, such as PIK3CD, BCL2, and CYCS, were confirmed through Western blot analysis. To evaluate apoptosis, we measured mitochondrial membrane potential and reactive oxygen species (ROS) levels, and to this end, employed molecular docking to analyze the binding affinity of active compounds toward PIK3CD.To functionally validate the role of PIK3CD, apoptosis-related indicators (mitochondrial membrane potential and reactive oxygen species (ROS) levels) were measured in PIK3CD-knockdown cells. Both the growth and metastatic migration of SW480 cells were potently attenuated following extract administration. Immunoblotting analyses demonstrated diminished expression levels of PIK3CD and cytochrome c , concomitant with elevated B-cell lymphoma 2 protein abundance—molecular alterations consistent with the initiation of programmed cell death. Mitochondrial membrane potential decreased, and ROS levels increased following PIK3CD knockdown, further supporting apoptosis. Molecular docking showed strong binding affinity between PIK3CD and stigmasterol, a key component of the extract. The AKT/mTOR transduction machinery was also modulated, suggesting a multi-targeted anti-cancer mechanism. Iphigenia indica Kunth extract exerts anti-cancer effects on CRC cells by inhibiting PIK3CD and the regulation of the AKT/mTOR signaling, which induces apoptosis. Collectively, these findings suggest the extract's potential as a CRC therapeutic, warranting further in vivo confirmation.