Zhoushan Hospital, Zhejiang, China.
Integr Cancer Ther. 2012 Dec;11(4):337-53. doi: 10.1177/1534735410392579. Epub 2011 Mar 7.
Previous clinical studies have shown good efficacy of the traditional Chinese medicinal herbal water extract Chan-Yu-Bao-Yuan-Tang (CYBYT) in lung cancer patients. In this study, CYBYT's effects on proliferation and apoptosis of human lung adenocarcinoma cell line SPC-A-1 cultured in vitro were explored. An XTT assay, cell cycle analysis, Annexin V-FITC staining and Western blot were applied to identify the viability of cells, cell cycle arrest, stages of apoptosis, and signaling proteins, respectively. The results showed that CYBYT inhibited the growth of SPC-A-1 cells by reducing the cells in G0/G1 phase but increasing them in S phase in a concentration-dependent manner, and inducing apoptosis, whereas it had no significant inhibitory effects on the normal human IMR-90 fibroblasts. Furthermore, early and total induction of apoptosis was positively correlated with the concentration of CYBYT in SPC-A-1 cells, and the rate of total apoptosis was greater in the CYBYT 100 µg/mL and 50 µg/mL groups than that of the positive control 5-fluorouracil (5-Fu) group. Moreover, CYBYT upregulated bax, cleaved caspase-3 protein expression, downregulated bcl-2 protein expression, and released mitochondrial cytochrome c into the cytosol in a time- and concentration-dependent manner. Our findings indicated that CYBYT could significantly inhibit growth and induce apoptosis via the mitochondrial pathway in human lung adenocarcinoma cell line SPC-A-1.
先前的临床研究表明,中药草药水提取物蟾榆保元汤(CYBYT)在肺癌患者中具有良好的疗效。在这项研究中,我们探讨了 CYBYT 对体外培养的人肺腺癌细胞系 SPC-A-1 增殖和凋亡的影响。应用 XTT 检测法、细胞周期分析、Annexin V-FITC 染色和 Western blot 分别鉴定细胞活力、细胞周期阻滞、凋亡阶段和信号蛋白。结果表明,CYBYT 呈浓度依赖性地减少 SPC-A-1 细胞的 G0/G1 期细胞数而增加 S 期细胞数,从而抑制细胞生长,诱导细胞凋亡,但对正常人类 IMR-90 成纤维细胞无明显抑制作用。此外,SPC-A-1 细胞中早期和总凋亡的诱导与 CYBYT 的浓度呈正相关,100 µg/mL 和 50 µg/mL CYBYT 组的总凋亡率大于阳性对照 5-氟尿嘧啶(5-Fu)组。此外,CYBYT 呈时间和浓度依赖性地上调 bax、cleaved caspase-3 蛋白表达,下调 bcl-2 蛋白表达,并将线粒体细胞色素 c 释放到细胞质中。我们的研究结果表明,CYBYT 可通过线粒体途径显著抑制人肺腺癌细胞系 SPC-A-1 的生长并诱导其凋亡。