淫羊藿苷促进白血病K562细胞凋亡的研究

Icaritin promotes the apoptosis of leukemia K562 cells

目的探讨淫羊藿苷对人慢性粒细胞白血病K562细胞的作用及其机制。方法将对数生长期K562细胞分成对照组和淫羊藿苷处理组。对照组细胞常规培养，淫羊藿苷处理组加入8μmol／L淫羊藿苷培养。应用四甲基偶氮唑盐（MTT）法检测K562细胞增殖，流式细胞术检测细胞凋亡，反转录聚合酶链反应（RT－PCR）检测细胞p85、AktmRNA的表达，Westernblot检测p85、Akt、p-p85、D—Akt、cleavage—caspase-3、caspase．3蛋白表达的变化。结果与对照组比较，淫羊藿苷处理组K562细胞增殖率明显下降（P〈0．05），K562细胞凋亡率和p85、Akt、cleavage—caspase－3蛋白表达均明显升高（均P〈0．05），p85mRNA、AktmRNA、caspase-3蛋白表达水平差异均无统计学意义（均P〉0．05）。结论淫羊藿苷能抑制K562细胞增殖，促进其凋亡，其作用机制可能是通过激活P13K—Akt信号转导通路实现的。

Objective To investigate the effect and mechanism of the icaritin on the human cronic myeloid leukemia K562 cells. Methods The K562 cells at logarithmic growth phase were divided into the control group and the icaritin group. The cells in the control group were normally treated and the cells in the icaritin group were incubated with 8 μmol/L icaritin. Methyl thiazolyl tetrazolium （MTT） method and flow eytometry were used to examine the proliferation and apoptotic changes in the two groups after incubation for 72 h, respectively. Gene expression of p85 and Akt were detected by RT-PCR. The protein changes of p85, Akt, p-p85, p-Akt eleavage-caspase-3 and caspase-3 were detected by Western blot. Results Compared with the control group, the proliferation rate of K562 cells in the icaritin group was significantly decreased （P 〈 0.05）, however the apoptotic rate of K562 cells and the expressions of p-p85, p-Akt, cleavage-caspase-3 in the icaritin group were significantly increased （all P 〈 0.05）, but the expressions of p85 mRNA, Akt mRNA and easpase-3 protein had no difference （all P 〉 0.05）. Conclusion Icaritin could induce the proliferation and promote the apoptosis of K562 cell, and its mechanism may be achieved through activating the PI3K-Akt signal transduction pathway.