二硫代氨基甲酸吡咯烷诱导人肝癌细胞凋亡的铜离子依赖性机制

目的：测定二硫代氨基甲酸吡咯烷（PDTC）处理对人肝癌细胞Hep3B凋亡的诱导作用及Cu^2 对这种作用的影响。方法：运用细胞增殖力、膜损伤、DNA片断化、细胞周期DNA含量测定分析PDTC对人肝癌细胞株Hep3B细胞的增殖抑制及凋亡诱导作用及Cu^2 在其中发挥的作用。结果：PDTC处理后，细胞增殖能力显著下降。乳酸脱氢酶（LDH）分析显示细胞膜并没有被破坏，DNA凝胶电泳及细胞周期DNA含量测定发现明显的凋亡特征性DNA片断及亚G1峰，显示细胞增殖能力的降低源于细胞凋亡的发生。低浓度Cu^2 显著加强了PDTC的细胞增殖抑制及凋亡诱导作用，而Cu^2 络合剂Bathocuproine disulfonate(BCS)显著抑制PDTC的两种作用。结论：PDTC铜离子依赖性地抑制人肝癌细胞增殖并诱导细胞凋亡。

Cu2+-dependent mechanism of pyrrolidine dithiocarbamate on human hepatoma cells apoptosis

Objective:To examine the effects of pyrrolidine dithiocarbamate (PDTC) on cells apoptosis and the function of Cu 2+ on this effects in human hepatoma Hep3B cells.Methods:Cells viability, membrane damage, DNA ladder and DNA content histograms were studied.Results:After being treated with PDTC, the proliferation of human hepatoma cells decreased remarkably, but cells membrane was not destroyed indicated by Lactate dehydrogenase (LDH) assay, and both DNA fragmentation on gel electrophoresis and sub-G1 DNA peak in DNA content histograms were clearly shown. This indicated that the loss of cells viability was resulted from the apoptosis induced by PDTC. Cu 2+ enhanced the function of PDTC on the induction of apoptosis, whereas bathocuproine disulfonate (BCS), a chelator of Cu 2+, obviously attenuated the induction of apoptosis.Conclusion:These results indicate that PDTC induces apoptosis of Hep3B cells via a Cu 2+-dependent manner.