三氮唑席夫碱衍生物对肝癌细胞裂亡的影响

目的:研究3-吡啶-3-基-4-[(4-甲氧基-苯亚甲基)氨基]-5-甲硫基-1,2,4-三唑(LH-38)对肝癌细胞BEL-7402裂亡的作用。方法:BEL-7402细胞常规培养于RPMI-1640培养液中,细胞生长至对数生长期加LH-38(终浓度分别为1×10-4mol/L和1×10-5mol/L),连续培养48 h或72 h。四甲基偶氮唑蓝(MTT)比色法检测细胞增殖,荧光染料Hoechst33258和PI联染检测细胞死亡,免疫细胞化学法检测激活型Caspase-3表达。结果:LH-38抑制BEL-7402细胞增殖并呈浓度依赖关系,IC50为3.0×10-4mol/L;1×10-5mol/L浓度的LH-38处理细胞72 h,镜下可见多倍体细胞明显增多,可见微核或多核细胞染色体自发性的凝集,有丝分裂异常,存活或死亡的多核或单核巨细胞同时存在,存活细胞的激活型Caspase-3表达阴性;1×10-4mol/L浓度LH-38处理细胞48 h,可明显诱导细胞凋亡。结论:不同浓度LH-38可以引起人肝癌细胞BEL-7402细胞裂亡或细胞凋亡,即两种不同形式的细胞死亡。

Effect of triazole schiff bases derivatives on mitotic catastrophe of hepatocarcinoma1 cell

Objective: To study the effect of 3-pyridin-3-yl-4-[(4-hydroxy-3-methoxy-benzylidene)amino]-5-methylsulfanyl-4H-[1,2,4] triazole(LH-38) on induction of mitotic catastrophe of hepatocarcinoma BEL-7402 cells.Methods: Human hepatocarcinoma cells of the line BEL-7402 were routinely cultured in RPMI-1640 Medium supplemented with 10% heat-inactivated fetal calf serum.Subconfluent cell cultures were treated with LH-38 at a final concentration of 1×10-4mol/L and 1×10-5mol/L for 48h or 72 h.The growth and proliferation of BEL-7402 cells were analyzed with MTT assay.The co-staining with Hoechst 33258 and propidium iodide were used to observe the cell death.The protein expressions of active caspase-3 in BEL-7402 cells were observed by immunohistochemical staining.Results: The cell proliferation was inhibited by LH-38 in dose dependent manners and the IC50 was 3.0×10-4mol/L.Treatment with LH-38(1×10-5mol/L) for 72 h increased the proportion of polyploid cells in BEL-7402 cells significantly and the spontaneous chromosome condensation and abnormal mitosis were observed in the cells with micronuclei and nuclear segments.The living or dead multinucleated and mononucleated giant cell could be observed simultaneously,and the protein expressions of active caspase-3 in temporarily viable cells were negative.1×10-4mol/L LH-38 also induced apoptosis of BEL-7402.Conclusion: Different doses of LH-38 may contribute to the induction of two distinct modes of cell death in BEL-7402 hepatocarcinoma cells,either apoptosis or cell death through mitotic catastrophe.