汉防己甲素抑制肝癌细胞增生的作用

Inhibitory mechanism on proliferation of hepatocellular carcinoma cell line by tetrandrine

目的:研究汉防己甲素(tetrandrine,TTD)对肝癌细胞的作用及其机制。方法:采用MTT比色法观察了TTD对肝癌细胞增生的影响;采用流式细胞仪法观察了TTD对肝癌细胞内活性氧的影响;采用琼脂凝胶电泳法观察了DNA片断化的梯形条带。结果:10 and 20μmol/L TTD作用于肝癌细胞系24,48,72h后其增生率分别为90.1±1.0％ and 77.5±2.0％,70.2±2.9％ and 56.6±1.6％,61.6±2.0％ and 47.2±1.9％(F=40.025,P<0.001);0、10和20μmol/L TTD作用于肝癌细胞系2h后O_2^-为35％、36.6％、63.2％;H_2O_2为24.5％、40.5％、84.6％。48h后20μmol/L TTD组肝癌细胞出现典型的凋亡表现。结论:TTD能够通过诱导肝癌细胞内活性氧的产生,而引起细胞凋亡,呈明显的剂量依赖性。

AIM: To study the mechanism of tetrandrine (TTD) on pro- liferation and apoptosis of hepatic cell carcinoma (HCC) cell line. METHODS: MTT colorimetry was used to evaluate the in- fluence of TTD on proliferation of cells. Flow cytometry was used to test reactive oxygen species (ROS) levels in cells. Electrophoresis in 1.5 % agarose gels was used to assess the DNA laddering. RESULTS: Proliferation rate of HCC cell line stimulated by 10 and 20 μmol/L TTD at 24 h, 48 h, and 72 h was 90.1±1.0 %, 77.5±2.0 %, 70.2±2.9 % and 56.6±1.6 %, 61.6±2.0 % and 47.2+1.9%, respectively (F=40.025, P<0.001). O_2^- and H_2O_2. generated by HCC cell lines incubated with 0, 10 and 2O μmol/L TTD for 2 hours were 35 and 24.5 %, 36.6 and 40.5 %, 63.2 and 84.6 %, respectively. Typical DNA ladder was observed after the cell line was incubated in 20 μmol/L TTD for 48 hours. CONCLUSION. TTD can induce the apoptosis of ROS of hepatic cell carcinoma ceil tine by generation of ROS in a dose dependent manner.