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目的:探讨瑞香狼毒药效组分Zp1111对人肝癌细胞的增殖抑制与诱导凋亡作用及机制。方法:MTT法比较瑞香狼毒药效组分Zp1111对体外培养的人来源肝癌细胞与正常肝细胞LO2抑制作用,采用流式细胞术检测用药前后BEL-7402细胞凋亡的变化并分析Zp1111对DNA周期分布的影响,荧光共振能量转移法检测Zp1111对细胞周期依赖性蛋白激酶CDK2的抑制作用。结果:Zp1111对体外培养的肝癌细胞HepG2、BEL-7402、SMMC-7721均有较好的抑制作用,IC50分别为37.75μg/ml、28.60μg/ml、29.22μg/ml,而对正常肝细胞LO2抑制作用不明显,在25μg/ml及以下的浓度对LO2细胞反而有一定促进作用。Zp1111能诱导BEL-7402细胞发生凋亡,一定药物浓度范围内凋亡率具有药物浓度依赖关系;Zp1111还能调控体外培养的BEL-7402细胞周期分布,显著下调S期的细胞比例,并且对CDK2具有较好的抑制作用。结论:瑞香狼毒药效组分Zp1111具有一定诱导凋亡作用,可使BEL-7402细胞周期阻滞在G1期,其作用机制可能与抑制CDK2有关。
Objective: To investigate the inhibitory effect and mechanism of anti-proliferation of human hepatic carcinoma cell line Zp1111 induced by Stellera chamaejasme L. on apoptosis. Method: MTT method is more active components Stellera chamaejasme Zp1111 source of cultured human hepatoma cells and normal liver cells LO2 inhibition of apoptosis using BEL-7402 cells before and after treatment and flow cytometry analysis of DNA Zp1111 Cycle distribution. Fluorescence resonance energy transfer assay was used to detect the inhibitory effect of Zp1111 on cell cycle-dependent protein kinase CDK2. Results: Zp1111 on cultured hepatoma cells HepG2, BEL-7402, SMMC-7721 had better inhibition, IC50 were 37.75μg / ml, 28.60μg / ml, 29.22μg / ml, while the normal liver cells LO2 Inhibition is not obvious, in the 25μg / ml and below the concentration of LO2 cells but have a certain role in promoting. Zp1111 capable of inducing apoptosis of BEL-7402 cells, within a certain range of drug concentrations apoptosis rate of drug having a concentration-dependent; Zp1111 also controls cultured BEL-7402 cell cycle distribution, significantly reduced the proportion of cells in S phase, and on CDK2 Has a good inhibitory effect. Conclusion: The active ingredient of Stellera chamaejasme L. Zp1111 can induce the apoptosis of BEL-7402 cells in G1 phase. Its mechanism may be related to the inhibition of CDK2.