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目的探讨青蒿琥酯经10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)介导的信号通路诱导人白血病K562细胞凋亡作用机制。方法将5个浓度(0,6.25,12.5,25,50μg·m L-1)浓度的青蒿琥酯作用K562细胞0-72h,通过噻唑蓝比色法观察其对K562细胞生长抑制作用;罗丹明123检测线粒体膜电位变化;荧光定量聚合酶链反应检测PTEN mRNA水平变化,试剂盒检测半胱天冬酶(Caspase)3/7活性;蛋白印迹法检测PTEN、蛋白激酶B(p-Akt)蛋白水平。结果青蒿琥酯能显著抑制K562细胞增殖,72 h最大抑制率为91.5%,与剂量和时间呈正相关。青蒿琥酯明显降低K562细胞线粒体膜电位,增加PTEN表达,抑制p-Akt表达,增强Caspase3/7蛋白活性。结论青蒿琥酯可能通过PTEN/Akt/Caspase通路诱导K562细胞凋亡。
Objective To investigate the mechanism of artesunate on the apoptosis of human leukemia K562 cells induced by phosphatase and tensin homolog (PTEN) mediated by chromosome 10. Methods K562 cells were treated with 5 artesunate concentrations of 0, 6.25, 12.5, 25 and 50 μg · m L-1 for 0-72 h. The growth inhibition of K562 cells was observed by thiazolyl blue colorimetry. Ming 123 was used to detect the change of mitochondrial membrane potential. The changes of PTEN mRNA level were detected by fluorescence quantitative polymerase chain reaction (RT-PCR), Caspase 3/7 activity was detected by kit, PTEN, p-Akt, Protein level. Results Artesunate significantly inhibited the proliferation of K562 cells. The maximum inhibition rate at 72 h was 91.5%, which was positively correlated with dose and time. Artesunate significantly decreased the mitochondrial membrane potential, increased the expression of PTEN, inhibited the expression of p-Akt, and enhanced the activity of Caspase 3/7 in K562 cells. Conclusion Artesunate may induce apoptosis of K562 cells through PTEN / Akt / Caspase pathway.