中药复方“松友饮”诱导肝癌细胞凋亡的实验研究

目的：研究中药复方“松友饮”对人肝癌细胞凋亡的影响及可能的分子机制，为临床应用提供理论依据。方法：采用流式细胞术、Caspase-3活性实验及RealTimePCR技术，观察2、4、8、10、20、30及40me,／mL“松友饮”（c1、c2、c3、c4、c5、c6及c7组）对体外培养的人肝癌细胞MHCC97H凋亡的影响，并探索可能的分子机制。结果：流式细胞仪检测结果显示，同对照组相比，干预组细胞凋亡率呈时间依赖性增加，“松友饮”（c4，10mg／mL）干预72h后MHCC97H细胞凋亡率增加超过3倍。干预48h后Caspase-3显著激活，Caspase-3活化呈时间依赖性。“松友饮”浓度增加，转移抑制基因1（Metastasissuppressor1，MTSS1）抑制效果越明显，“松友饮”（c4，10mg／mL）干预48h，MTSS1基因表达抑制率为39．0％。结论：“松友饮”诱导肝癌细胞凋亡可能与MTSS1基因表达抑制、Casoase-3活化相关。

Experimental Research on Apoptosis of Hepatocellular Carcinoma Induced by Chinese Herbal Compound Songyou Yin

Objective： To investigate the mechanism of Chinese herbal compound Songyou Yin inducing apoptosis in human hepatocellular carcinoma （HCC）, and to provide the experimental basis of Songyou Yin in treatment for HCC.Methods ： Flow cytometry （ FCM）, Caspase - 3 experiment, and Real - Time PCR were applied to test changes of HCC treated by Songyou Yin at final concentrations of 0 （ cO group, control） ,2,4,8,10,20,30,40 mg/mL （ cl, c2, c3, c4, c5, c6, and c7 groups, respectively） for 24,48,72 h, and the underlying mechanisms were explored. SAS 8.2 software was applied for statistical analysis, and the significance was defined as P 〈 0.05. Results：MHCC97H cells were sensitive to Songyou Yin. Songyou Yin （ c4,10 mg/mL） did not cause MHCC97H cell cycle arrest at GO/G1 until 72 h treatment after the release of synchronized cultures via FCM analysis, but the apoptosis rate of treated cells was higher than that in control in a time -dependent manner throughout the period of study. The in vitro apoptosis index by FCM analysis increased up to about threefold following 72 h of treatment by Songyou Yin （c4,10 mg/mL,P 〈0.05）. We observed that Songyou Yin caused a significant apoptosis increase in MHCC97H ceils, where longer treatment times were more effective. To examine the apoptosis common effectors via the measurement of Caspase -3 in vitro with nontoxic Songyou Yin treatment （c4,10 mg/mL） ,our data revealed that Caspase- 3 was evidently activated after 48 h of treatment, accorded with the inhibited effect on metastasis suppressor 1 （MTSS1） by inhibition rate of 39.0% （P 〈 0.05 ）. Conclusion： Nontoxic herbal compound extract Songyou Yin inducing HCC apoptosis may be correlated with activation of Caspase - 3 and the inhibition effect on MTSS1 expression.