摘要
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 induction of apoptosis of human liver cancer line SMMC7721 treated with electrochemical therapy(ECT,that is,direct current),and to provide the basis for clinical application of ECT.Methods The effect of ECT was investigated in vitro.The apoptosis and cell cycle arrest induced by ECT were determined by Annexin V-affinity assay,DNA content analysis and electron microscope.The effects of ECT on p53 and p21 WAF1/CIP1 protein expression were assessed by immunohistochemistry assay.The change in expression of p21 WAF1/CIP1 mRNA after ECT treatment was detected by RT-PCR assay.Results Of Annexin V-affinity assay display,the apoptosis rate of SMMC7721 cells was significantly increased after 24h and 48h of ECT tratment compared with the control(P<0.05).After 48h of ECT treatment,nuclear chromatin condensation of some cells was found under electron microscope.Sub-G1 cell population and ratio of S and G2/M cell population were increased by DNA content assay.On TR-PCR method and immunohistochemistry assay,it was found that the expression level of p21 WAF1/CIP1 mRNA and protein was enhanced.Conclusion The study shows that the ECT induces the apoptosis and cell cycle arrest of human liver cancer line SMMC7721.Enhancing the expression of p21 WAF1/CIP1 may be one of mechanisms.
出处
《中华理疗杂志》
2000年第1期37-40,共4页
Chinese Journal of Physical Therapy
