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目的 研究电化学治疗（ ＥＣＴ） 对人肝癌细胞系ＳＭＭＣ７７２１ 凋亡的诱导作用及其机制，为ＥＣＴ治疗的临床应用提供依据。方法 采用体外模型研究ＥＣＴ 的效应。以Ａｎｎｅｘｉｎ Ｖ 标记法、ＤＮＡ含量测定、电子显微镜等方法观察ＥＣＴ 对人肝癌细胞系ＳＭＭＣ７７２１ 凋亡的诱导作用及对细胞周期的阻滞效应。免疫组化法观察ＥＣＴ 对ｐ５３ 、ｐ２１ ＷＡＦ１／ＣＩＰ１ 蛋白的影响，ＲＴ－ ＰＣＲ 法观察ＥＣＴ对ｐ２１ ＷＡＦ１／ＣＩＰ１ ｍ ＲＮＡ表达水平的影响。结果 Ａｎｎｅｘｉｎ Ｖ 标记法显示，与对照组相比，ＥＣＴ作用后２４，４８ ｈ 细胞凋亡率明显增高（Ｐ＜ ０ ．０５） 。ＥＣＴ（５Ｖ２Ｃ） 作用后４８ ｈ，电镜下见部分细胞核染色质凝聚，ＤＮＡ 含量检测显示亚Ｇ１ 峰增高，Ｓ期及Ｇ２／ Ｍ 期细胞比例增高，ＲＴ－ ＰＣＲ、免疫组化法检测显示ｐ２１ ＷＡＦ１／ＣＩＰ１ｍＲＮＡ及蛋白表达水平升高。结论 ＥＣＴ 治疗能诱导人肝癌细胞ＳＭＭＣ７７２１ 凋亡及周期阻滞，上调ｐ２１ ＷＡＦ１／ＣＩＰ１ 的表达可能为其机制之一。
Objective To study the effect of electrochemical treatment (ECT) on the apoptosis of human hepatocellular carcinoma cell line SMMC7721 and its mechanism, and provide evidence for the clinical application of ECT. Methods The effect of ECT was studied in vitro. Annexin V labeling method, DNA content determination, electron microscopy and other methods ECT on human hepatocellular carcinoma cell line SMMC7721 apoptosis induction and cell cycle arrest effect. The effect of ECT on p53 and p21 WAF1 / CIP1 protein expression was observed by immunohistochemistry. The effect of ECT on the expression of p21 WAF1 / CIP1 m RNA was observed by RT-PCR. Results Annexin V labeling showed that compared with the control group, the apoptotic rate was significantly increased at 24 and 48 h after ECT treatment (P <0.05). 48h after ECT (5V2C), some nuclear chromatin condensation was observed under electron microscope. The content of sub-G1 was increased and the proportion of cells in S phase and G2 / M phase was increased by DNA content assay. RT-PCR and immunohistochemistry showed that p21 WAF1 / CIP1 mRNA and protein expression levels increased. Conclusion ECT treatment can induce the apoptosis and cell cycle arrest of human hepatocellular carcinoma cell line SMMC7721. Upregulating the expression of p21 WAF1 / CIP1 may be one of its mechanisms.