Furusawa S, Wu J, Fujimura T, Nakano S, Nemoto S, Takayanagi M, Sasaki K, Takayanagi Y
Department of Pharmacology and Toxicology, Cancer Research Institute, Tohoku College of Pharmacy, Sendai, Japan.
Methods Find Exp Clin Pharmacol. 1998 Mar;20(2):87-97. doi: 10.1358/mf.1998.20.2.485636.
Cepharanthine, a biscoclaurine alkaloid extracted from Stephania cepharantha Hayata was examined for a possible apoptosis-inducing effect in murine P388 doxorubicin-sensitive (P388/S) and -resistant (P388/DOX) cells. A significant increase in LDH release from cells was observed after P388/S and P388/DOX cells had been exposed to cepharanthine for 24 h. Cepharanthine (10 micrograms/ml) markedly induced apoptosis in resistant cells after 6 h and 24 h. By the means of agarose electrophoresis the DNA ladder was detected in cell lines treated with cepharanthine for 24 h. Cepharanthine (1-10 micrograms/ml) also induced the production of reactive oxygen species in P388/DOX cells, while no such cepharanthine-induced increase in reactive oxygen species was observed in P388/S cells. Flow cytometry analysis showed a high level of Fas-antigen expression in P388/DOX cells treated with cepharanthine. Furthermore, we found that the inhibitition of DNA and protein synthesis caused by cepharanthine (10 micrograms/ml) was more significant in resistant cells than in sensitive cells. Cepharanthine had no effect on the GSH content of P388/S and P388/DOX cells. Our experimental results suggest that cepharanthin can induce apoptosis both in P388/S and P388/DOX cells, especially in the latter. Apoptosis induced by cepharanthine may be implicated in the production of reactive oxygen species and Fas-antigen expression in tumor cells.
头花千金藤碱是从千金藤中提取的一种双苄基异喹啉生物碱，本研究检测了其对小鼠P388阿霉素敏感细胞（P388/S）和耐药细胞（P388/DOX）的凋亡诱导作用。P388/S和P388/DOX细胞经头花千金藤碱处理24小时后，细胞乳酸脱氢酶释放显著增加。头花千金藤碱（10微克/毫升）在6小时和24小时后可显著诱导耐药细胞凋亡。经琼脂糖凝胶电泳检测，用头花千金藤碱处理24小时的细胞系中出现了DNA梯状条带。头花千金藤碱（1 - 10微克/毫升）也可诱导P388/DOX细胞产生活性氧，而在P388/S细胞中未观察到头花千金藤碱诱导的活性氧增加。流式细胞术分析显示，用头花千金藤碱处理的P388/DOX细胞中Fas抗原表达水平较高。此外，我们发现头花千金藤碱（10微克/毫升）对DNA和蛋白质合成的抑制作用在耐药细胞中比在敏感细胞中更显著。头花千金藤碱对P388/S和P388/DOX细胞的谷胱甘肽含量没有影响。我们的实验结果表明，头花千金藤碱可诱导P388/S和P388/DOX细胞凋亡，尤其是后者。头花千金藤碱诱导的凋亡可能与肿瘤细胞中产生活性氧和Fas抗原表达有关。
