Kim Dae Won, Ahan Song Ho, Kim Tae Young
Department of Neurosurgery, School of Medicine, Wonkwang University, Iksan, Korea.
J Korean Neurosurg Soc. 2007 Nov;42(5):392-9. doi: 10.3340/jkns.2007.42.5.392. Epub 2007 Nov 20.
Arsenic trioxide (As(2)O(3)) 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 As(2)O(3) 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 As(2)O(3) and 75 microg/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 As(2)O(3) or berberine only.
Combined treatment of As(2)O(3) 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.
三氧化二砷（As₂O₃）在传统中药中作为抗癌剂已使用了数千年，黄连素是一种异喹啉生物碱，具有显著的抗菌活性。我们研究了As₂O₃和黄连素联合对人神经母细胞瘤（HNB）SH - SY5Y细胞的体外抗癌作用，并阐明其潜在的分子机制。
用2 microM As₂O₃和75 microg/ml黄连素处理HNB SH - SY5Y细胞，通过MTT法、DAPI染色、琼脂糖凝胶电泳、流式细胞术分析和蛋白质免疫印迹分析评估细胞存活、细胞死亡机制以及协同细胞毒性作用。
两种药物联合处理也显著降低了细胞活力。两种药物的细胞毒性表现为凋亡，其特征为染色质浓缩、DNA片段化和线粒体膜电位丧失。凋亡细胞毒性伴随着caspase - 3蛋白酶的激活以及Bcl - 2、Bid和Bcl - x/L表达的降低。此外，与仅用As₂O₃或黄连素处理的细胞相比，两种药物联合处理的细胞还显示细胞内活性氧水平和脂质过氧化显著增加。
As₂O₃与黄连素联合处理诱导HNB SH - SY5Y细胞凋亡信号通路的激活。这些结果表明低浓度的两种化疗药物联合治疗有可能以最小的副作用提高对癌细胞的细胞毒性作用。
