Department of Biochemistry, Kangwon National University, Chuncheon 200-701, Republic of Korea.
J Ethnopharmacol. 2010 Mar 24;128(2):342-51. doi: 10.1016/j.jep.2010.01.049. Epub 2010 Feb 4.
Cordyceps is a parasitic fungus and has long been used as a traditional Chinese medicine to treat illnesses, promote longevity, increase athletic power, and relieve exhaustion and cancer. In this study, we reveal the mechanisms underlying apoptosis induced by Cordyceps pruinosa butanol fraction (CPBF) in the human cervical adenocarcinoma cell line, HeLa.
Proliferation and apoptosis of cells were examined by MTT assay, DNA fragmentation, phosphatidyl serine distribution assay, Western blot analysis, and immunocytochemistry. To determine the association between CPBF related apoptosis and ROS, electron spin resonance (ESR) trapping experiments were used.
CPBF inhibited proliferation and induced apoptosis in HeLa cells in a dose-dependent manner using a MTT assay, DNA fragmentation, and a phosphatidyl serine distribution assay. Western blot analysis showed that apoptosis in HeLa cells was caspase-3- and -9-dependent. Proteolytic cleavage of PARP and the release of cytochrome c from the mitochondria into the cytosol were significantly increased and the Bcl-2/Bax protein ratio was decreased. Apoptosis induced by CPBF was not prevented by various antioxidants.
These results indicate that apoptotic effects of CPBF on HeLa cells are mediated by mitochondria-dependent death-signaling pathway independent of reactive oxygen species, suggesting that CPBF might be effective as an anti-proliferative agent for cancer.
冬虫夏草是一种寄生真菌,长期以来一直被用作中药来治疗疾病、延年益寿、增强运动能力以及缓解疲劳和癌症。在这项研究中,我们揭示了蛹虫草正丁醇级分(CPBF)诱导人宫颈腺癌 HeLa 细胞凋亡的机制。
通过 MTT 检测、DNA 片段化、磷脂酰丝氨酸分布分析、Western blot 分析和免疫细胞化学检测来检测细胞的增殖和凋亡。为了确定 CPBF 相关凋亡与 ROS 之间的关系,使用电子自旋共振(ESR)捕获实验。
CPBF 通过 MTT 检测、DNA 片段化和磷脂酰丝氨酸分布分析,以剂量依赖的方式抑制 HeLa 细胞的增殖并诱导其凋亡。Western blot 分析表明,HeLa 细胞的凋亡依赖于 caspase-3 和 -9。PARP 的蛋白水解裂解以及细胞色素 c 从线粒体释放到细胞质中明显增加,Bcl-2/Bax 蛋白比例降低。各种抗氧化剂不能预防 CPBF 诱导的细胞凋亡。
这些结果表明,CPBF 对 HeLa 细胞的凋亡作用是通过线粒体依赖性死亡信号通路介导的,与活性氧无关,提示 CPBF 可能作为一种有效的抗癌增殖剂。