Zhang Yongkui, Zeng Fang, Liu XiaoGuang, Li YunCheng, Zhou JiHang, Huang YanYan, Wang Yekai, Zhou ShiQuan, Zhu WangYu, Shu ErYun, Zhou Guoer, Chen Gang
Joint Laboratory of Immunogenomics, Zhoushan Hospital-Beijing Institute of Genomics, Chinese Academy of Sciences, Zhoushan, China.
Xenobiotica. 2011 Jul;41(7):593-602. doi: 10.3109/00498254.2011.565818. Epub 2011 Mar 29.
Previous clinical studies have shown efficacy and safety of the traditional Chinese medicinal herb extract Chan-Yu-Bao-Yuan-Tang (CYBYT) in lung cancer patients. The effects of CYBYT on proliferation and apoptosis in the non-small cell lung cancer cell line NCI-H460 and the small cell lung cancer cell line NCI-H446 were investigated in vitro. CYBYT significantly induced antiproliferative effects of NCI-H460 and NCI-H446 cells in a concentration- and time-dependent manner. The IC(50) values in NCI-H460 cells were 94.37 µg/mL (24 h) and 20.89 µg/mL (48 h), whereas in NCI-H446 cells IC(50) values were 214.72 µg/mL (24 h) and 114.58 µg/mL (48 h). Annexin V-FITC/PI staining showed CYBYT could significantly induce apoptosis in NCI-H460 and NCI-H446 cells, and the total apoptosis rates were positively correlated with the concentration and time of CYBYT treatment. Furthermore, treatment with the broad-spectrum caspase inhibitor (z-VAD-fmk) effectively protected NCI-H460 and NCI-H446 cells against CYBYT-triggered apoptosis. The apoptotic processes involved were a marked decrease in antiapoptotic protein Bcl-2 and an increase in proapoptotic protein Bax. The release of mitochondrial cytochrome c into the cytosol was also observed, which, in turn, resulted in the activation of caspase-9 and caspase-3. CYBYT exerts antiproliferative and growth inhibition effects on NCI-H460 and NCI-H446 cells through the mitochondrial caspase-dependent cell death pathway.
先前的临床研究已表明，中药提取物蟾鱼保元汤（CYBYT）对肺癌患者具有疗效和安全性。本研究在体外考察了CYBYT对非小细胞肺癌细胞系NCI-H460和小细胞肺癌细胞系NCI-H446增殖和凋亡的影响。CYBYT能以浓度和时间依赖性方式显著诱导NCI-H460和NCI-H446细胞的抗增殖作用。NCI-H460细胞的IC(50)值在24小时时为94.37μg/mL，48小时时为20.89μg/mL；而在NCI-H446细胞中，IC(50)值在24小时时为214.72μg/mL，48小时时为114.58μg/mL。膜联蛋白V-FITC/PI染色显示CYBYT能显著诱导NCI-H460和NCI-H446细胞凋亡，总凋亡率与CYBYT处理的浓度和时间呈正相关。此外，用广谱半胱天冬酶抑制剂（z-VAD-fmk）处理可有效保护NCI-H460和NCI-H446细胞免受CYBYT诱导的凋亡。所涉及的凋亡过程包括抗凋亡蛋白Bcl-2显著减少和促凋亡蛋白Bax增加。还观察到线粒体细胞色素c释放到细胞质中，进而导致半胱天冬酶-9和半胱天冬酶-3活化。CYBYT通过线粒体半胱天冬酶依赖性细胞死亡途径对NCI-H460和NCI-H446细胞发挥抗增殖和生长抑制作用。
