Berberine, a natural alkaloid sensitizes human hepatocarcinoma to ionizing radiation by blocking autophagy and cell cycle arrest resulting in senescence
| Autor: | Gautham Ramesh, Satish Rao Bola Sadashiva, Shubhankar Das |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Radiation-Sensitizing Agents
Carcinoma Hepatocellular Cell cycle checkpoint Berberine Pharmaceutical Science Mitochondria Liver Mitochondrion 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Sequestosome-1 Protein Autophagy Humans MTT assay Mitotic catastrophe Cellular Senescence Cell Proliferation 030304 developmental biology Pharmacology 0303 health sciences Cell growth Liver Neoplasms Hep G2 Cells Cell biology G2 Phase Cell Cycle Checkpoints Oxidative Stress chemistry 030220 oncology & carcinogenesis Microtubule-Associated Proteins Intracellular |
| Zdroj: | Journal of Pharmacy and Pharmacology. 72:1893-1908 |
| ISSN: | 2042-7158 0022-3573 |
| Popis: | ObjectiveTo study the radiosensitizing potential of Berberine and the underlying mechanism in human hepatocarcinoma (HepG2) cells.MethodsHepG2 cells were challenged with X-rays in combination with Berberine treatment and several in vitro assays were performed. Alteration in cell viability was determined by MTT assay. Changes in intracellular ROS levels, mitochondrial membrane potential/mass, intracellular acidic vesicular organelles as well as cell cycle arrest and apoptotic cell death were analysed by flow cytometry. Induction of autophagy was assessed by staining the cells with Monodansylcadaverine/Lysotracker red dyes and immunoblotting for LC3I/II and p62 proteins. Phase-contrast/fluorescence microscopy was employed to study mitotic catastrophe and senescence. Cellular senescence was confirmed by immunoblotting for p21 levels and ELISA for Interleukin-6.Key findingsX-rays + Berberine had a synergistic effect in reducing cell proliferation accompanied by a robust G2/M arrest. Berberine-mediated radiosensitization was associated with elevated levels of LC3II and p62 suggesting blocked autophagy that was followed by mitotic catastrophe and senescence. Treatment of cells with X-rays + Berberine resulted in increased oxidative stress, hyperpolarized mitochondria with increased mitochondrial mass and reduced ATP levels.ConclusionsThe study expands the understanding of the pharmacological properties of Berberine and its applicability as a radiosensitizer towards treating liver cancer. |
| Databáze: | OpenAIRE |
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