Hu Sheng, Chen Shi-min, Li Xiao-kuan, Qin Rui, Mei Zhi-nan
Institute of Materia Media, South-Central University for Nationalities, Wuhan, China.
Acta Pharmacol Sin. 2007 Aug;28(8):1215-23. doi: 10.1111/j.1745-7254.2007.00606.x.
To investigate the antihepatocellular carcinoma effects of chi-shen extract (CSE) from the water-soluble compounds of Salvia miltiorrhiza and Paeoniae radix.
The effect of CSE on the growth of HepG2 cells (hepatocellular carcinoma cell line) was studied by 3-(4,5)-2,5-diphenyltetrazolium bromide assay. Apoptosis were detected through acridine orange (AO) and ethylene dibromide (EB) staining and DNA fragmentation assay. The effect of CSE on the cell cycle of HepG2 cells was studied by the propidium iodide staining method. The activation of caspases-3, -8 and -9 was examined by immunoassay kits. The transcription of the Bcl-2 family and p53 was detected by RT-PCR.
Our data revealed that CSE strongly induced HepG2 cell death in a dose- and time-dependent manner. CSE-induced cell death was considered to be apoptotic by observing the typical apoptotic morphological change by AO/EB staining and DNA fragmentation assay. The induction of HepG2 cell death was caused by an induction of apoptosis for the sub-G1 proportion increase, the downregulation of Bcl-2, the upregulation of Bax and p53, and the activation of the caspases-3 and -9 pathways.
These results clearly demonstrated that CSE was able to inhibit the proliferation of HepG2 cells and cause apoptosis. Moreover, the anticancer effects of CSE were related to the Bcl-2 family pathway and the activation of caspases-3 and -9 in HepG2 cells.
研究丹参和芍药水溶性成分的赤参提取物(CSE)对肝癌细胞的抗癌作用。
采用3-(4,5)-2,5-二苯基四氮唑溴盐法研究CSE对HepG2细胞(肝癌细胞系)生长的影响。通过吖啶橙(AO)和溴化乙锭(EB)染色及DNA片段化分析检测细胞凋亡情况。采用碘化丙啶染色法研究CSE对HepG2细胞周期的影响。使用免疫分析试剂盒检测半胱天冬酶-3、-8和-9的激活情况。通过逆转录聚合酶链反应(RT-PCR)检测Bcl-2家族和p53的转录情况。
我们的数据显示,CSE以剂量和时间依赖性方式强烈诱导HepG2细胞死亡。通过AO/EB染色和DNA片段化分析观察到典型的凋亡形态变化,认为CSE诱导的细胞死亡为凋亡性死亡。HepG2细胞死亡的诱导是由于亚G1期比例增加、Bcl-2下调、Bax和p53上调以及半胱天冬酶-3和-9通路激活导致的凋亡诱导。
这些结果清楚地表明,CSE能够抑制HepG2细胞的增殖并导致细胞凋亡。此外,CSE的抗癌作用与HepG2细胞中的Bcl-2家族通路以及半胱天冬酶-3和-9的激活有关。