Background: Cancer remains a major global health challenge, characterized by uncontrolled cell growth and metastasis. Colorectal cancer (CRC) is one of the most common malignancies worldwide, highlighting the urgent need for effective therapeutic strategies. This study aimed to evaluate the therapeutic potential of hispolon in HT-29 human colorectal adenocarcinoma cells, with a focus on its effects on cell viability, oxidative stress markers, and apoptosis.
Objectives: To investigate the apoptotic and cytotoxic effects of hispolon on the human colorectal adenocarcinoma cell line (HT-29).
Materials and Methods: The HT-29 cell line was obtained from the National Centre for Cell Science (NCCS), Pune, India. Hispolon (98% purity) was procured from Santa Cruz Biotechnology (sc-221726), Dallas, Texas, USA. HT-29 cells were treated with varying concentrations of hispolon (10–100 µM). Cell viability was assessed using the MTT assay. Lipid peroxidation was evaluated by measuring malondialdehyde (MDA) formation. Antioxidant enzyme activities, including superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx), were analyzed, and reduced glutathione (GSH) levels were quantified. Reactive oxygen species (ROS) generation was assessed using a fluorescent probe.Apoptosis was evaluated using acridine orange/ethidium bromide (AO/EtBr) staining.
Results and Discussion: Hispolon treatment exhibited dose-dependent cytotoxic effects (10–100 µM) on HT-29 cells. A significant reduction in cell viability was observed, with an IC?? value of 48 µM, as determined by the MTT assay. Increased lipid peroxidation and elevated MDA levels indicated the induction of oxidative stress. Hispolon treatment altered antioxidant enzyme activities, with increased SOD and catalase activities and decreased GPx activity. Additionally, GSH levels were significantly reduced in hispolon-treated cells. A marked increase in ROS generation was observed following hispolon exposure. AO/EtBr staining demonstrated a significant increase in apoptotic cell populations in treated cells.
Conclusion: The findings suggest that hispolon exerts potent anticancer effects on HT-29 cells by inducing oxidative stress, modulating antioxidant defense mechanisms, and promoting apoptosis. The observed dose-dependent cytotoxicity, increased lipid peroxidation, enhanced ROS generation, and altered antioxidant enzyme activities highlight the potential of hispolon as a therapeutic agent for colorectal cancer. Further studies are warranted to elucidate the underlying molecular mechanisms and explore its clinical applicability.
Acknowledgements: The authors gratefully acknowledge financial support from the Chief Minister’s Research Grant (CMRG), Government of Tamilnadu(Grant No. CMRG2023BBC03022).