苦参碱抑制KG1a细胞生长及提高自然杀伤细胞对其杀伤敏感性的研究

Matrine inhibits growth of KG1a cells and enhances the cytotoxicity of natural killer cells

目的:观察苦参碱(matrine)对人急性髓系白血病细胞株KG1a的生长抑制作用,及苦参碱作用前后自然杀伤(natural killer,NK)细胞对KG1a细胞杀伤敏感性的影响。方法:采用CCK-8法和锥虫蓝染色法检测苦参碱对KG1a细胞的生长抑制作用及细胞存活率,FCM检测苦参碱作用前后KG1a细胞周期的变化及表面NK细胞活化性受体(natural-killer group 2,member D,NKG2D)配体(MICA/B、ULBP1、ULBP2和ULBP3)和人类白细胞抗原(human leukocyte antigen,HLA)Ⅰ类分子的表达,乳酸脱氢酶释放法检测苦参碱作用前后NK细胞对KG1a细胞杀伤敏感性的影响。结果:苦参碱能抑制KG1a细胞的生长,随着药物浓度的升高和作用时间的延长,细胞的生长抑制率逐渐升高、存活率逐渐下降;苦参碱作用后细胞发生G1/S期阻滞;细胞表面NKG2D各配体表达率均明显升高,与作用前相比差异有统计学意义(P<0.05),HLA-Ⅰ类分子表达率无明显变化(P>0.05);当效靶比分别为5:1、10:1和20:1时,苦参碱作用前后NK细胞对KG1a细胞的杀伤敏感性差异均有统计学意义(P<0.05)。结论:苦参碱可抑制KG1a细胞的生长并改变细胞周期,上调KG1a细胞表面NKG2D各配体的表达率,增强NK细胞对其的杀伤敏感性。

Objective：To observe the inhibitory effects of matrine on growth of human acute myelocytic leukemia KG1a cell line and the enhancing effects of matrine on cytotoxicity of natural killer （NK） cells against KG1a cells. Methods：The inhibitory effects of matrine on growth and survival rate of KG1a cells were measured by CCK-8 assay and Trypan blue staining method, respectively. The cell cycle distribution and expression of natural killer group 2 member D （NKG2D） ligands （MICA/B, ULBP1, ULBP2, and ULBP3） and human leukocyte antigen （HLA） class Ⅰ molecules on KG1a cells were assayed by flow cytometery before and after matrine treatment. Cytotoxicity of NK cells isolated from 6 healthy volunteers against KG1a cells was analyzed by LDH releasing assay at different effector-to-target cell ratios （E∶T）. Results：Matrine inhibited the growth and survival rate of KG1a cells in a time-and concentration-dependent manner. The growth inhibition rate gradually increased and survival rate gradually decreased with the increase in drug concentration and extension of treatment period. KG1a cells was blocked at G1/S phase and the expression ratio of NKG2D ligands as eleva-ted after matrine treatment. The difference was significant （P〈0.05）. There was no significant difference in expression of HLA classⅠmolecules before and after matrine treatment （P〈0.05）. The cytotoxicity of NK cells was significantly enhanced at 5∶1, 10∶1, 20∶1of E∶T ratio after matrine treatment （P〈0.05）. Conclusion：Matrine inhibits the growth of KG1a cells, changes cell cycle distribution, up-regulated the expression of NKG2D ligands, and enhances the sensitivity of KG1a cells to the cytotoxicity of NK cells.