Hsu Shu-Chun, Kuo Chao-Lin, Lin Jing-Pin, Lee Jau-Hong, Lin Chin-Chung, Su Chin-Cheng, Lin Hui-Ju, Chung Jing-Gung
Graduate Institute of Chinese Pharmaceutical Sciences, China Medical University, Taichung 404, Taiwan, ROC.
Anticancer Res. 2007 Jul-Aug;27(4B):2415-25.
In this study, the effects of 95% ethanol extracts of Euchresta formosana radix (EFR) on the cell cycle and apoptosis in human hepatocellular carcinoma (HCC) Hep3B cells were investigated. The results indicated that EFR decreased DNA synthesis and viable Hep3B cell numbers in a concentration-dependent manner. EFR induced a p21- and p27-dependent cell cycle arrest in S-phase and apoptosis of the Hep3B cells. The induction of apoptosis by EFR treatment was also confirmed by DAPI staining. EFR inhibited cyclin-dependent kinase (CDK)-1 and -2 expression and decreased cyclin B1 and E levels, resulting in S-phase arrest. EFR induced reactive oxygen species (ROS) production followed by endoplasmic reticulum (ER) stress that was based on the increase of GADD153 and GRP78 which led to the release of Ca2+ in the Hep3B cells. The EFR-promoted apoptosis was associated with increasing activation of caspases 3, 7, and 9 and enhanced poly(ADP-ribose) polymerase cleavage and increased expression of p21(CIP1/WAF1), p27(KIP1), Bax and Bad. Furthermore, the levels of Bcl-xl decreased after EFR treatment. Alteration of these key anti- and pro-apoptotic proteins could contribute to the increase in p53-independent apoptosis that was observed in the Hep3B cells.
在本研究中，考察了台湾山豆根95%乙醇提取物（EFR）对人肝癌Hep3B细胞的细胞周期及凋亡的影响。结果表明，EFR以浓度依赖性方式降低DNA合成及存活的Hep3B细胞数量。EFR诱导Hep3B细胞在S期发生依赖p21和p27的细胞周期阻滞及凋亡。DAPI染色也证实了EFR处理诱导的凋亡。EFR抑制细胞周期蛋白依赖性激酶（CDK）-1和-2的表达，降低细胞周期蛋白B1和E的水平，导致S期阻滞。EFR诱导活性氧（ROS）生成，随后引发内质网（ER）应激，这是基于Hep3B细胞中GADD153和GRP78增加导致Ca2+释放。EFR促进的凋亡与半胱天冬酶3、7和9的激活增加、聚（ADP-核糖）聚合酶切割增强以及p21（CIP1/WAF1）、p27（KIP1）、Bax和Bad表达增加有关。此外，EFR处理后Bcl-xl水平降低。这些关键的抗凋亡和促凋亡蛋白的改变可能导致在Hep3B细胞中观察到的p53非依赖性凋亡增加。
