EFFECT OF TETRADIUM RUTICARPUM AND ITS BIOACTIVE COMPOUNDS ON INTRINSIC APOPTOTIC SIGNALLING IN ORAL SQUAMOUS CELL CARCINOMA (OSCC) CELLS IN VITRO: ROLE OF WNT/β-CATENIN PATHWAYS

Background: A major global health concern, oral cancer is becoming more common due to a combination of environmental and hereditary factors. Oral cancer is primarily caused by dysregulation of the Wnt/β-catenin signaling pathway, which promotes cell proliferation and inhibits apoptosis. As such, it is an important target for therapeutic intervention. Tetradium ruticarpum extracts may be able to modulate the Wnt/β-catenin pathway to restore apoptotic mechanisms and stop the growth of tumors. Materials and method: The DPPH radical scavenging assay was used to measure antioxidant activity, while the albumin denaturation inhibition method was used to measure anti-inflammatory qualities. In a CO2 incubator, human oral cancer KB cells were grown in DMEM containing medium supplemented with 10% FBS and 1% antibiotics. Using the MTT and trypan blue tests, cytotoxicity was investigated. Using Real-Time PCR, the gene expression of apoptotic markers was examined. To investigate compound-protein interactions, molecular docking studies were carried out using PyRx and Biovia Discovery Studio. One-way ANOVA was used to evaluate the data (p<0.05). Results: Results of Preliminary biochemical analysis showed that rutaecarpine significantly improved DPPH radical scavenging activity and inhibited albumin denaturation suggesting the strong antioxidant and anti-inflammatory potential (p<0.001). Treatment of rutaecarpine to oral cancer cells (KB) resulted in dose-dependent antiproliferative activity in both 20 and 40µM concentrations (p<0.002). Futher, gene expression anlaysis by q-RT-PCR registered the compound’s ability to inhibit the over expression of Wnt/Beta catenin signaling molecules (β-catenin, Bcl-2 and Wnt mRNA). Molecular docking anlyais also authenticated the results of cell line model by showing the strong binding intercations with the target molecules (β-catenin, Bcl-2 and Wnt). Conclusion: The potential of rutaecarpine as an anticancer treatment is highlighted in this work by its capacity to inhibit inflammatory processes, decrease cell viability, and modify apoptotic and wnt/ β-catenin signaling pathways in human oral cancer cells. Future research is made possible by its molecular docking and antioxidant results, which further support its therapeutic promise against oral cancer.