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目的:探讨manumycin对人髓系白血病细胞株K562细胞的增殖抑制作用,为其治疗白血病提供理论依据。方法:实验设阴性对照组(K562加含10%胎牛血清的IMDM培养液)和manumycin组(终浓度分别为2、4、8及16μmol·L-1),分别作用24、48和72h,镜下观察manumycin作用后K562细胞的生长情况;MTT法检测K562细胞的生长抑制率;24h后,用Western blotting方法检测对照组及4、16μmol·L-1manumycin组K562细胞中survivin蛋白的表达。结果:镜下观察显示,对照组细胞生长状态良好,密度均一,大小一致,轮廓清晰,折光性强。与对照组比较,8μmol·L-1 manumycin组细胞明显变小,密度稀疏,大小不均一,细胞轮廓欠清晰,折光性差;16μmol·L-1 manumycin组细胞密度明显减少,大小不等,轮廓不清,有细胞碎片。MTT结果显示,随着manumycin浓度增加和作用时间延长,生长抑制率逐渐增加,与对照组比较,差异有统计学意义(P<0.05或P<0.01),且存在时间-剂量-效应关系。Western blotting结果显示,与对照组比较,16μmol·L-1 manumycin作用K562细胞24h以后细胞内survivin蛋白含量下降。结论:manumycin通过下调K562细胞survivin蛋白的表达,促进细胞凋亡,抑制肿瘤细胞的生长,从而达到治疗白血病的目的。
Objective: To investigate the inhibitory effect of manumycin on the proliferation of human myeloid leukemia cell line K562 and to provide a theoretical basis for the treatment of leukemia. Methods: The negative control group (K562 plus IMDM medium containing 10% fetal bovine serum) and manumycin group (2,4,8 and 16 μmol·L-1, respectively) The growth of K562 cells was observed under microscope. The growth inhibition rate of K562 cells was detected by MTT assay. After treated with 4, 16μmol·L-1manumycin for 24 hours, the expression of survivin in K562 cells was detected by Western blotting. Results: Microscopic observation showed that the control group grew well, with uniform density, uniform size, clear outline and strong refraction. Compared with the control group, the cells in 8 μmol·L-1 manumycin group were significantly smaller, the density was sparse, the size was not uniform, the cell outline was not clear and the refraction was poor. The cell density in 16 μmol·L-1 manumycin group was significantly reduced, Clear, there are cell debris. The results of MTT showed that with the increase of manumycin concentration and the prolongation of action time, the growth inhibition rate gradually increased. Compared with the control group, the difference was statistically significant (P <0.05 or P <0.01), and there was a time-dose-effect relationship. Western blotting results showed that compared with the control group, K562 cells treated with 16μmol·L-1 manumycin decreased the protein expression of survivin in 24h. Conclusion: Manumycin can inhibit the growth of tumor cells by down-regulating the expression of survivin protein in K562 cells, so as to achieve the goal of treating leukemia.