达沙替尼联合氟达拉滨对慢粒K562细胞的抑制作用研究

目的探讨达沙替尼联合氟达拉滨对慢粒K562细胞的抑制作用。方法选取慢粒K562细胞株进行研究，采用MTT法分别测定单独使用达沙替尼和氟达拉滨对慢粒K562细胞的抑制率，以及达沙替尼联合氟达拉滨对诱导K562细胞的抑制率。根据金氏方程计算2种药物联合的抑制率及凋亡情况的协同作用及治疗效果。金氏公式为:q=D1+2/(D1+D2-D1×D2)，q表示2种药物联合作用的抑制率，D1和D2是单独用药作用的抑制率。当q值＞1.15表示为协同作用。经过不同浓度的达沙替尼(1，5，10 μg/L)和氟达拉滨(1，2.5，5 ng/L)单独处理后或联合处理(1 μg/L达沙替尼+1 ng/L氟达拉滨)，(5 μg/L达沙替尼+2.5 ng/L氟达拉滨)，(10 μg/L达沙替尼+5 ng/L氟达拉滨)24 h后，K562细胞的增殖受到明显的抑制，且达沙替尼和氟达拉滨具有协同效应。结果在相同的时间范围内，氟达拉滨和达沙替尼对K562细胞的抑制作用呈剂量依赖性，由于5 μg/L达沙替尼和2.5 ng/L氟达拉滨均能明显抑制K562细胞的增殖作用，因此在后续的实验过程中，选择该浓度作为细胞处理的终浓度。实验组和对照组的抑制率，差异均有统计学意义(t＝39.998，P<0.05)。达沙替尼联合氟达拉滨存在协同抑制慢粒K562细胞的作用(q＞1.15，P＜0.05)；低浓度(1 μg/L)达沙替尼对慢粒K562细胞p-BCR/ABL水平的下调作用(31.8％±1.9％)明显优于高浓度(10 ng/L)氟达拉滨(15.2％±2.1％)，联合药物更明显下调慢粒K562细胞p-BCR/ABL水平的表达(49.8％±1.1％)，差异具有统计学意义(t＝6.754，P＜0.05)。达沙替尼联合氟达拉滨能改善白血病，提高凋亡细胞的数量。结论达沙替尼联合氟达拉滨作用于白血病慢粒K562细胞具有协同抑制作用，加速慢粒K562细胞的凋亡，具有重要的临床意义。

Study on Inhibition of Dasatinib and Fludarabine to CML K562 Cells

ObjectiveTo study the inhibition of Dasatinib and fludarabine on CML K562 cells．MethodsCML K562 cells were analyzed，the MTT method was used to determine the inhibition of CML K562 cells，dasatinib and fludarabine,respectively,and the inhibition rate of dasatinib joint fludarabine to CML K562 cells．According to kimformula,calculated the synergistic effect and the therapeutic effect of the inhibition rate and apoptosis of 2 kinds of drug combinations ［Kim formulais:q=D1+2/(D1+D2-D1×D2)］．q means the inhibition rate of two drug combination，D1 and D2 inhibition rate of the medicine effect．When the q value is greater than 1.15 was expressed as synergy，according to the guinness book of equation proved the inhibition rate and apoptosis of synergy and the treatment effect of two drug combination．Determined by MTT method to detect the results showed that after different concentrations for Dasatinib (1，5，10 μg/L) and fluorine (1，2.5，5 ng/L) after treatment alone or combined processing (1 μg/L Dasatinib＋1 ng/L fludarabine)，(5 μg/L Dasatinib+2.5 ng/L fludarabine)，(10 μg/L Dasatinib+5 ng/L fludarabine) after 24 h，inhibition of the proliferation of K562 cells were obvious，and Dasatinib and fluorine had a synergistic effect．ResultsAt the same time within the scope of fluorine dara marina and for sand for inhibition of K562 cells was dose dependent，because mu 5 g/L for sand for shore and 2.5 ng/L fluorine dara could significantly inhibits the proliferation of K562 cells，so in subsequent experiments，chose the concentration as the final concentration cell processing．The inhibition rate of the experimental group and control group，all had statistical significance (t=39.998，P< 0.05)．For sand for joint fluorine dara marina exist K562 cells of synergistic inhibition CML (q>1.15，P<0.05)；Low concentrationmu (1 g/L) for sand for K562 cells of CML p-lower BCR/ABL level role (31.8%±1.9%) high concentration (10 ng/L) was obviously better than the fluorine dara marina (15.2%±2.1%)，combination of drugs more apparent K562 cells by CML p-the expression of BCR/ABL levels (49.8%±1.1%)，statistically significant difference (t=6.754，P<0.05)．For sand for joint fluorine dara marina can improve leukemia，increase the number of apoptotic cells．ConclusionDasatinib and fludarabine effect on CML K562 cellshave synergistic inhibition effect，accelerate the apoptosis of CML K562 cells，has important clinical significance．