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目的探讨藤黄酸对白血病细胞系K562增殖、凋亡、细胞周期的影响,并观察藤黄酸对hERG钾通道蛋白的调控作用。方法K562细胞以不同浓度藤黄酸(0.125~8.0μmol/L)处理0~72 h后,MTT法观察藤黄酸对K562细胞生长抑制的情况,Annexin-V/PI双标法及透射电镜检测细胞凋亡,PI单染法检测细胞周期分布,Western blotting、RT-PCR法分别检测藤黄酸对K562细胞内hERG通道的调控作用。结果藤黄酸能明显抑制K562细胞增殖,具有时间-剂量依赖性,其24 h的IC50为(2.637±0.208)μmol/L。此外,藤黄酸以浓度依赖性方式诱导K562细胞凋亡,并伴随明显的凋亡细胞形态学改变,而藤黄酸的凋亡诱导效应可能与其诱导K562细胞周期阻滞于G0/G1期有关。hERG钾通道蛋白在人白血病K562细胞中表达量较高,藤黄酸对hERG钾通道蛋白及其表达水平均有不同程度的抑制作用,该抑制作用呈明显的量效关系(P<0.01)。结论藤黄酸可通过下调hERG钾通道蛋白的表达发挥较强的抗白血病效应,hERG钾通道有望成为白血病诊治的新靶标。
Objective To investigate the effect of gambogic acid on the proliferation, apoptosis and cell cycle of leukemia cell line K562, and to observe the regulation effect of gambogic acid on hERG potassium channel protein. Methods K562 cells were treated with different concentrations of gambogic acid (0.125-8.0 μmol/L) for 0-72 h. MTT assay was used to observe the growth inhibition of K562 cells by Annexin-V/PI double labeling and transmission electron microscopy. Apoptosis was detected by PI staining. Western acid and RT-PCR were used to detect the regulation of hERG channels in K562 cells. RESULTS Gambogic acid significantly inhibited the proliferation of K562 cells in a time- and dose-dependent manner. The IC50 at 24 h was (2.637±0.208) μmol/L. In addition, gambogic acid induces apoptosis of K562 cells in a concentration-dependent manner, with apoptotic cell morphology changes, and the apoptosis-inducing effect of gambogic acid may be related to its induction of K562 cell cycle arrest in G0/G1 phase. . The hERG potassium channel protein was highly expressed in human leukemia K562 cells. Gambogic acid inhibited the expression of hERG potassium channel protein and its expression in different degrees. The inhibitory effect was significant (P<0.01). Conclusion Gambogic acid can exert a strong anti-leukemia effect by down-regulating the expression of hERG potassium channel protein. The hERG potassium channel is expected to become a new target for the diagnosis and treatment of leukemia.