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目的探讨大麻素WIN55,212-2(WIN)对肝癌细胞BEL-7402增殖和凋亡的影响,并对其机制进行初步研究。方法将细胞分成对照组和WIN处理组,处理细胞后,利用MTT法分析WIN对BEL-7402细胞增殖的影响;DAPI染色分析各组细胞中细胞核的形态学改变;流式细胞术检测各实验组的细胞凋亡率;利用荧光探针JC-1检测细胞线粒体膜电位的变化;Western blot检测WIN处理细胞后Bcl-2蛋白的表达;分光光度法检测Caspase-3、-8和-9的活性。结果WIN抑制了BEL-7402细胞增殖并诱导其凋亡,两者都具有剂量依赖性;WIN处理细胞后,Caspase-3、-8和-9活性总体上呈时间依赖性升高,而且Bcl-2蛋白表达下降。结论 WIN能抑制BEL-7402细胞增殖和诱导其凋亡,WIN诱导的BEL-7402细胞凋亡可能与线粒体途径和死亡受体途径都有关。这些结果为应用WIN来治疗肝癌奠定了一个基础。
Objective To investigate the effects of cannabinoids WIN55 and 212-2 (WIN) on the proliferation and apoptosis of hepatocellular carcinoma cell line BEL-7402, and to study its mechanism. Methods The cells were divided into the control group and the WIN treatment group. After treatment of the cells, the effect of WIN on the proliferation of BEL-7402 cells was analyzed by MTT assay. The morphological changes of nuclei in each group were analyzed by DAPI staining. Flow cytometry The apoptosis rate of cells was detected by fluorescent probe JC-1. The expression of Bcl-2 protein was detected by Western blot and the activity of Caspase-3, -8 and -9 was detected by spectrophotometry . Results WIN inhibited the proliferation and induced apoptosis of BEL-7402 cells in a dose-dependent manner. After treated with WIN, the activities of Caspase-3, -8 and -9 were increased in a time-dependent manner in general, and Bcl- 2 protein expression decreased. Conclusion WIN can inhibit the proliferation and induce the apoptosis of BEL-7402 cells. WIN-induced apoptosis of BEL-7402 cells may be related to both mitochondrial and death receptor pathways. These results lay a foundation for the treatment of liver cancer with WIN.