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- Title
β-sitosterol induces reactive oxygen species-mediated apoptosis in human hepatocellular carcinoma cell line.
- Authors
Ditty, Mary J.; Ezhilarasan, Devaraj
- Abstract
Objective: It is of interest to investigate the anti-proliferative effect of β-sitosterol (BS) on human hepatocellular carcinoma (HepG2) cell line. Materials and Methods: β-sitosterol treatments (0.6 and 1.2 mM/ml) were done in HepG2 and after 24 hr, cell viability was evaluated by MTT assay. Reactive oxygen species (ROS) accumulating potential of BS was assessed by dichloro-dihydrofluorescein diacetate staining. Morphology related to apoptosis was investigated by acridine orange and ethidium bromide dual staining. Cytochrome c and caspase 3 expressions were evaluated by immunofluorescence and western blot analyses. Results: β-sitosterol induced cytotoxicity (p<0.001) and intracellular ROS in HepG2 cells in a dose-dependent manner. BS treatments accumulated induced intracellular ROS accumulation which led to membrane damage and mitochondrial toxicity. At the molecular level, BS treatments induced cytochrome c release from mitochondria and enhanced the protein expressions (p<0.05 vs 0.6 mM/ml and p<0.001 vs 1.2 mM/ml) of both caspase 3 and cleaved caspase 3. Conclusion: β-sitosterol induced ROS accumulation which plays a critical role in apoptosis via the intrinsic pathway in HepG2 cells. The present investigation paves the way for further in vivo studies.
- Subjects
REACTIVE oxygen species; HEPATOCELLULAR carcinoma; CELL lines; MITOCHONDRIAL proteins; CYTOCHROME c; APOPTOSIS
- Publication
Avicenna Journal of Phytomedicine, 2021, Vol 11, Issue 6, p541
- ISSN
2228-7930
- Publication type
Article
- DOI
10.22038/AJP.2021.17746