正肝方药物血清诱导人肝癌细胞分化的体外实验研究

目的：探讨正肝方对Bel-7402人肝癌细胞分化的诱导作用，及对其端粒酶活性的影响。方法：正肝方水煎液灌喂正常大鼠，采血制备药物血清。以正常鼠血清为阴性对照，维甲酸(RA)为阳性对照，将药血清温育Bel-7402人肝癌细胞。噻唑蓝(MTT)比色法测定细胞增殖，图像分析系统测定细胞核质比，放射免疫法测定甲胎蛋白(AFP)含量，动态比色法测定γ-谷氨酰转肽酶(γ-GT)和碱性磷酸酶(ALP)活性，多聚酶链反应-酶联免疫吸附法(PCR—ELISA法)检测细胞端粒酶活性。结果：正肝方药物血清可抑制人肝癌细胞的增殖，显著降低细胞核质比，抑制细胞分泌AFP，降低细胞γ-GT活性，升高ALP活性，并有显著抑制人肝癌细胞端粒酶活性的作用。结论：正肝方具有诱导人肝癌细胞分化的作用，其主要机理可能是抑制了Bel-7402细胞端粒酶活性。

Experimental Study on Differentiation of Human Hepatocarcinoma Cells Induced by Zhengganfang Drug Serum in Vitro

OBJECTIVE: To study the effect of Zhengganfang (ZGF) drug serum in inducing differentiation of Bel-7402 human hepatocarcinoma cell (HHC) and its influence of the telomerase activity. METHODS: Rats were fed with ZGF decoction to prepare the drug-serum of ZGF, and the drug-serum was used for Bel-7402 HHC culture. Serum of normal rat, was taken as negative control and retinoic acid (RA) as the positive control. Cell proliferation was detected by MTT colorimetric assay, nucleocytoplasm ratio by image assay, alpha-fetoprotein (AFP) by RIA, alkaline phosphatase (ALP) activity and gamma-glutamyl transpeptidase (gamma-GT) by dynamic colorimetric assay and telomerase activity of Bel-7402 cell by PCR-ELISA. RESULTS: Drug-serum of ZGF could inhibit the proliferation of Bel-7402 cell, markedly reduce the nucleocytoplasm ratio and the secretion of AFP, decrease gamma-GT activity, increase ALP activity and suppress the telomerase activity of Bel-7402 cell. CONCLUSION: ZGF can induce the differentiation of Bel-7402 HHC, the main mechanism maybe through the suppression of telomerase activity.