清毒饮、养正片影响白血病K562细胞增殖周期、细胞凋亡的实验研究

目的:观察清毒饮、养正片诱导人白血病细胞株K562细胞凋亡及对细胞增殖周期的作用;探索其时效关系,为临床提供选择最佳用药时机的实验基础.方法:制备含清毒饮、养正片药物血清,建立相应的培养体系,加入含药小鼠血清,培养K562细胞株;分别于培养12、24、36、48、72h收集细胞,固定、染色、上流式细胞仪进行细胞周期、细胞凋亡的检测.结果:人白血病K562细胞株存在有自然凋亡,正常小鼠血清无诱导该细胞凋亡的作用;清毒饮、阿糖胞苷具有诱导细胞凋亡的作用,且随着培养时间的延长凋亡率愈加明显.清毒饮对K562细胞周期的影响,还表现出明显的时间依赖性;随着培养时间的延长,当细胞出现明显凋亡时,细胞株S期显著下降,G0/G1期升高,提示清毒饮和清毒加养正合剂都可引起K562细胞株的G0/G1期阻滞,对凋亡的敏感性增强.结论:清毒饮能抑制K562细胞增殖、促进细胞分化、诱导细胞凋亡,提示该中药复方具有从分子水平抗肿瘤细胞增殖的效应,这也是中药复方协同化疗药物治疗白血病在增效减毒方面的关键机制;清毒饮对K562细胞发生细胞毒作用,是通过抑制其细胞周期S期、将其阻滞于G1期,并通过促进K562细胞凋亡实现的,其作用强度与时间呈密切正相关.

Experimental Study of QING-DU Decoction and YANG-ZHENG Tablet on the Growth,Cell Cycle and Apoptosis of K562 Cells

Objective:To observe the effects on Qing-Du Decoction,Yang-Zheng Tablet on various concentrations on the growth,cell cycle and apoptosis of K562 cells using flow cytometry.And to investigate the relationships of effectiveness for a given period of time in order to provide the basis of experiment for clinic with thebest time and dose of treatment.Methods:The serum of normal and contain-drugswere made first.Human acute cell line K562 cells were selected as target cells which cultured in the variant culturing systems contain serum of mouse.The culturing cells were selected on 12,24,36,and 48 and 72 hours,which were fixed,counted,colored and measured by flow cytometry.Results:There was natural apoptosis of K562 and the vacant serum group could not induce apoptosis.The Qing-Du decoction group had the action of inducing apoptosis and with the elongation of the time the apoptosis rate can increase.The effects of Qing-Du decoction cell cycle of K562 cells were expressed on time-dependent manner.When the apoptosis appeared,the S phrase of cell cycle could decrease and the G 0/G 1 phrase increasedwhich indicate the Qing-Du decoction and Qing-Yang decoction could arrest cells in the G 0/G 1 and increase the apoptosis rate of the K562 cells.Conclusion:Qing-Du decoction could arrest the growth of the K562 cells,increase the differentiation and induce the apoptosis which indicate Chinese herbal compound formulas had the effects of anti-proliferation of tumor cell on molecular biology level.We could conclude that the effects of Qing-Du decoction was restraint of the DNA synthesize in S phase,such as obstruct the metabolism of protein and choke back the fission of tumor cells.