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目的:测定二乙基二硫代氨基甲酸钠(DDC)对人肝癌细胞增殖、再分化与凋亡的作用。方法:采用细胞表面电荷、生化变化、软琼脂细胞生长、单细胞电泳、电子超微结构和流式细胞DNA片断分析等 指标测定细胞分化和凋亡。结果:用DDC 3mmol/L处理后,肝癌细胞的生长和分裂指数显著下降,增殖抑制率达52.4％。与恶化有关的指标显著减轻,如细胞表面电荷明显降低,电泳率从1.6降低到0.8μm·s~(-1)·V~(-1)·cm~(-1),甲胎蛋白由314降为95μg/g(protein),γ-谷氨酰转氨酶活性由0.9降到0.14U/g(protein)。与分化相关的酪氨酸-α-酮戊二酸转氨酶活性显著上升,由11.6升高到36μmol/g(protein),细胞克隆形成力降低95.6％。同时,处理细胞出现了凋亡小体、不贴壁细胞以及其它凋亡特征,且处理细胞DNA发生片断化,流式细胞分析显示处理组42.9％的DNA发生断裂。结论:DDC能够抑制人肝癌细胞增殖,并诱导细胞分化和凋亡。
Objective: To determine the effect of sodium diethyldithiocarbamate (DDC) on the proliferation, differentiation and apoptosis of human hepatocellular carcinoma cells. METHODS: Cell differentiation and apoptosis were determined using cell surface charge, biochemical changes, soft agar cell growth, single cell electrophoresis, electron ultrastructure, and flow cytometric DNA fragmentation analysis. RESULTS: After treatment with DDC 3mmol/L, the growth and division index of hepatoma cells were significantly decreased, and the proliferation inhibition rate was 52.4%. The indicators related to deterioration were significantly reduced, such as a significant decrease in cell surface charge, the electrophoresis rate decreased from 1.6 to 0.8 μm·s -1 -1 ·V -1 ·cm -1, and alpha-fetoprotein was controlled by 314 When reduced to 95 μg/g (protein), γ-glutamyl transaminase activity decreased from 0.9 to 0.14 U/g (protein). The tyrosine-alpha-ketoglutarate transaminase activity associated with differentiation increased significantly from 11.6 to 36 [mu]mol/g (protein), and the cell colony formation power decreased by 95.6%. At the same time, apoptotic bodies, non-adherent cells, and other apoptotic features appeared in the treated cells, and the DNA of the treated cells was fragmented. Flow cytometric analysis showed that 42.9% of the DNA in the treated group was broken. Conclusion: DDC can inhibit the proliferation of human hepatoma cells and induce cell differentiation and apoptosis.