紫草素诱导白血病细胞K562凋亡的研究

Study on the apoptotic induction of shikonin on human leukemia K562 cells

目的探讨紫草素对人白血病K562细胞的诱导凋亡作用。方法MTT显色法检测紫草素对K562细胞的增殖抑制作用;形态学观察分析DNA琼脂糖凝胶电泳;AnnexinV/PI双标记法检测紫草素对K562细胞的凋亡效应;Caspase检测K562细胞凋亡通路。结果紫草素0.25~8μg/ml浓度即能明显抑制K562细胞的增殖,并具有时间和浓度的依赖性。1μg/ml紫草素作用48h后的K562细胞出现凋亡细胞的特征。1~4μg/ml浓度紫草素诱导K562细胞凋亡具有明显的量效关系,2μg/ml紫草素在0~72h内诱导K562细胞凋亡具有明显的时效关系;DNA凝胶电泳呈现细胞凋亡的典型梯形条带;紫草素诱导K562细胞凋亡经历了Caspase-3、Caspase-6和Caspase-9的激活。结论紫草素能有效地诱导K562细胞凋亡,并主要通过线粒体凋亡通路。

Objective To evaluate the apoptotic induction of shikonin on hmnan leukemia K562 cells. Methods MTr colorimetric assay was used to examine the growth inhibition of shikonin on K562 cells. The apoptosis of K562 cells was detected by cell morphological observation, DNA agarose gel electrophoresis and Annexin V/PI double labeling. The apoptosis pathway was tested by activation of caspase. Results Shikonin significantly inhibited the growth of K562 cells in a concentration - depedent and time - dependent manner at die range of 0.25 to 8ug/ml. The K562 cells treated with 1.0ug/ml shikonin for 48 hours showed morphological characteristics of apoptotic cells; The K562 cells treated with shikonin of different concentration （1.0, 2.0, 4. 0ug/ml） for 48 hours ; with 2.0μg/ml shikonin for 24, 48 and 72 hours; induction of apoptosis was associated with exposure time and concentration. There was typical ladder strap in DNA gel electrophoresis. Shikonin induced K562 cells apoptosis involved in the activation of caspase - 3, caspase - 6 and caspase - 9. Conclusion Shikonin can induce apoptosis of hmnan leukemia K562 cells, and the key is mitochondrial apoptosis pathways.