萝卜硫素诱导白血病细胞K562凋亡及其与化疗药协同作用的实验研究

Experimental Study of Sulforaphane Effect on Cell Apoptosis of K562 and Its Synergistic Effect with Chemotherapy Drugs

目的:探讨萝卜硫素诱导K562白血病细胞作用及其与化疗药的协同作用。方法:琼脂半固体集落培养法和MTT法检测不同浓度的萝卜硫素对K562细胞增殖的影响;倒置显微镜观察萝卜硫素对细胞形态的影响,Annexin V-FITC/PI双标记法检测细胞凋亡情况;Western blot检测凋亡调控蛋白PARP的活性;观察萝卜硫素分别和高三尖杉酯碱、阿糖胞苷联合的抑瘤作用,用药物相互作用指数评价药物协同作用。结果:萝卜硫素能够抑制白血病K562的增殖,呈剂量依赖性;萝卜硫素引起了K562细胞形态的明显改变,显著提高了细胞的早期凋亡率,增加了促凋亡蛋白PARP的表达水平;萝卜硫素与化疗药物高三尖杉酯碱和阿糖胞苷存在明显的协同作用,其协同系数分别为0.64和0.66。结论:萝卜硫素可以诱导K562细胞发生凋亡,增加化疗药物的敏感性,具有良好的临床应用前景。

Objective ：To investigate the apoptosis inducing effects of Sulforaphane （SF） in human chronic myeloid leu- kemia cell K562 and its synergistic effect with chemotherapy drugs, homoharringtonine （Horn） and cytosine arabinoside （Ara -c）. Methods：KS62 cells were firstly treated with different concentrations of SF. Then, the inhibitory effect of SF was detected by MTY and semisolid agar culture assay;morphological changes of cells were observed by inverted microscope;cell apoptosis were detected by Annexin V - FITC/PI staining. Western blot was carried out to determine the expression of ap- optosis related proteins PARP. Finally,the synergistic effects of SF with Horn and Ara - c were determined respectively. Re- sults：SF significantly inhibited the proliferation,induced cell apoptosis and morphological changes of K562 cells. Further- more,SF significantly increased the activities of pro - apoptotic proteins PARP. SF also increased sensitivity of K562 to- wards Hom and Ara - c. The combination indexes of SF with Horn and Ara - c were 0.64 and 0.66, respectively. Conclu- sions ： Our resuhs showed that SF could induce cell apoptosis of human chronic myeloid leukemia cell and increase the sensitivity to chemotherapy drugs,indicating potential clinical applications for chronic myeloid leukemia patients.