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Journal of Korean Neurosurgical Society 2007;42(5): 392-399.
doi: https://doi.org/10.3340/jkns.2007.42.5.392
Enhancement of Arsenic Trioxide (As2O3)-Mediated Apoptosis Using Berberine in Human Neuroblastoma SH-SY5Y Cells.
Dae Won Kim, Song Ho Ahan, Tae Young Kim
Department of Neurosurgery, School of Medicine, Wonkwang University, Iksan, Korea. tykim@wonkwang.ac.kr
Arsenic trioxide (As2O3) has been used as an anticancer agent in traditional Chinese medicine for thousand years and berberine is an isoquinoline alkaloid present that has indicated significant antimicrobial activity. We have examined the combined anticancer effects of As2O3 and berberine against the human neuroblastoma (HNB) SH-SY5Y cells in vitro, and to elucidate underlying molecular mechanism.
HNB SH-SY5Y cells were treated with 2 microM As2O3 and 75 microgram/ml berberine, and their survival, cell death mechanism as well as synergistic cytotoxic effects were estimated by using MTT assay, DAPI staining, agarose gel electrophoresis, flow cytometric analysis, and western blot analysis.
The combined treatment of two drugs also markedly decreased cell viability. The cytotoxic effects of two drugs were revealed as apoptosis characterized by chromatin condensation, DNA fragmentation, and the loss of mitochondrial membrane potential. The apoptotic cytotoxicity was accompanied by activation of caspase-3 protease as well as decreased the expression of Bcl-2, Bid, and Bcl-x/L. In addition, the cells treated with combination of two drugs also showed significantly increased intracellular reactive oxygen species levels and lipid peroxidation compared to cells As2O3 or berberine only.
Combined treatment of As2O3 with berberine induced activation of apoptotic signaling pathways in HNB SH-SY5Y cells. These results suggest that the possibility of the combined treatment of two chemotherapeutic agents with low concentration improving cytotoxic effect for cancer cells with minimal side effects.
Key Words: Human neuroblastoma SH-SY5Y cells; Arsenic trioxide; Berberine; Apoptosis
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