去氢木香内酯对慢性粒细胞白血病K562细胞增殖的影响及其机制

Effect of dehydrocostus lactone on the proliferation of chronic myeloid leukemia K562 cells and its mechanism

目的探讨去氢木香内酯对慢性粒细胞白血病细胞株K562细胞增殖的影响及其机制。方法用不同浓度去氢木香内酯作用于对数生长期的K562细胞,采用瑞特-吉姆萨染色观察细胞形态,CCK-8法检测细胞增殖情况,流式细胞术检测细胞周期、凋亡情况及细胞表面分化抗原CD14和CD11b表达,蛋白质印迹法检测JAK-STAT通路、细胞凋亡及细胞周期相关蛋白表达。结果不同浓度(4.0、6.0、8.0、10.0、12.0μmol/L)去氢木香内酯作用24h均能抑制K562细胞增殖,与对照组比较,差异有统计学意义(F=109.510,P<0.05)。5.0、10.0μmol/L去氢木香内酯作用24h后,K562细胞凋亡率分别为(16.1±3.8)%、(29.6±4.3)%,较对照组的(3.1±0.5)%升高(F=83.255,P<0.05)。5.0、10.0μmol/L去氢木香内酯作用24h后,K562细胞的G2/M期细胞比例分别为(17.0±3.2)%、(28.8±3.9)%,较对照组的(9.1±2.3)%升高(F=161.598,P<0.05);S期细胞比例分别为(48.1±3.9)%、(61.0±5.4)%,较对照组的(39.6±3.6)%升高(F=192.356,P<0.05)。2.5、5.0μmol/L去氢木香内酯作用72h后,K562细胞的CD14表达率分别为(28.6±3.9)%、(41.1±4.4)%,较对照组的(3.1±0.5)%升高(F=132.811,P<0.05);K562细胞的CD11b表达率分别为(42.4±5.0)%、(61.2±5.7)%,较对照组的(4.2±1.1)%升高(F=179.553,P<0.05)。去氢木香内酯能够降低JAK2、STAT5、cyclinE、CDK2、cyclinA、CDC25C、cyclinB1、CDK1及bcl-2蛋白表达,上调p21及bax蛋白的表达。结论去氢木香内酯能够抑制慢性粒细胞白血病K562细胞增殖,可能是通过细胞周期阻滞、诱导凋亡及分化而实现的。

Objective To explore the effect of dehydrocostus lactone on the proliferation of chronic myeloid leukemia cell line K562 and its mechanism.Methods K562 cells in logarithmic growth phase were treated with different concentrations of dehydrocostus lactone.Cell morphology was observed by Wright-Giemsa staining.Cell proliferation was detected by CCK-8 method.Cell cycle,apoptosis and the expressions of cell surface differentiation antigen CD14 and CD11b were detected by flow cytometry.JAK-STAT pathway and the expressions of apoptosis and cell cycle related proteins were detected by Western blot.Results Compared with the control group,the proliferation of K562 cells could be inhibited after treatment with different concentrations (4.0,6.0,8.0,10.0 and 12.0 μmol/L) of dehydrocostus lactone for 24 h,and the difference was statistically significant (F = 109.510,P < 0.05).After treatment with 5.0 and 10.0 μmol/L dehydrocostus lactone for 24 h,the apoptosis rates of K562 cells were (16.1±3.8)% and (29.6±4.3)%,which were higher than that of the control group [(3.1±0.5)%](F = 83.255,P < 0.05).After treatment with 5.0 and 10.0 μmol/L dehydrocostus lactone for 24 h,the proportion of G2/M phase cells in K562 cells were (17.0±3.2)% and (28.8±3.9)%,which were higher than that of the control group [(9.1±2.3)%](F = 161.598,P < 0.05);the proportion of S phase cells in K562 cells were (48.1±3.9)% and (61.0±5.4)%,which were higher than that of the control group [(39.6±3.6)%](F = 192.356,P < 0.05).After treatment with 2.5 and 5.0 μmol/L dehydrocostus lactone for 72 h,the expression rates of CD14 in K562 cells were (28.6±3.9)% and (41.1±4.4)%,which were higher than that in the control group [(3.1±0.5)%](F = 132.811,P < 0.05);the expression rates of CD11b in K562 cells were (42.4±5.0)% and (61.2±5.7)%,which were higher than that in the control group [(4.2±1.1)%](F = 179.553,P < 0.05).Dehydrocostus lactone could decrease the expressions of JAK2,STAT5,cyclin E,CDK2,cyclin A,CDC25C,cyclin B1,CDK1 and bcl-2 proteins,and up-regulate the expressions of p21 and bax proteins.Conclusion Dehydrocostus lactone can inhibit the proliferation of chronic myeloid leukemia K562 cells,which may be achieved by cell cycle arrest,induction of apoptosis and differentiation.