阿糖胞苷对姜黄素诱导的人急性髓系白血病细胞自噬的影响  

Effect of cytarabine on curcumin-induced autophagy in human acute myeloid leukemia cells

作　　者：马翠[1] 白天鸽 陈雅琳 魏虹[1] MA Cui;BAI Tian-ge;CHEN Ya-lin;WEI Hong(Department of Histology and Embryology, College of Medicine of Shihezi University, Shihezi 832002 , China)

机构地区：[1]石河子大学医学院组织胚胎学教研室

出　　处：《中国病理生理杂志》2019年第8期1409-1415,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81460024)

摘　　要：目的:观察化疗药物阿糖胞苷作用下姜黄素是否诱导人急性髓系白血病KG1a细胞产生自噬及可能的机制。方法:体外培养KG1a细胞,透射电镜观察不用药物处理后细胞超微结构的变化;用吖啶橙染色检测细胞内酸性自噬泡的变化;MTT法检测细胞活力变化;流式细胞术检测细胞周期的变化;RT-qPCR和Western blot检测自噬相关因子beclin-1和LC3 mRNA及蛋白的表达。结果:姜黄素对KG1a细胞的活力有明显抑制作用(P<0.05),且呈剂量依赖关系,姜黄素联合阿糖胞苷组细胞活力抑制率明显高于单药组,且两组均高于对照组(P<0.01)。电镜结果显示姜黄素能诱导细胞自噬小体的产生,阿糖胞苷能使姜黄素诱导的自噬小体增加。吖啶橙染色发现联合用药组能提高姜黄素诱导的细胞自噬,细胞内酸性自噬泡及含有自噬泡的细胞数均增加。细胞周期结果显示细胞主要被阻滞在G0/G1期。RT-qPCR结果显示联合用药组beclin-1和LC3的mRNA表达水平均比单药组及对照组明显升高(P<0.01);Western blot结果显示联合用药组比单药组beclin-1的蛋白表达明显上调(P<0.05),LC3-II/LC3-I的比值升高。结论:姜黄素能抑制KG1a细胞活力并诱导细胞发生自噬,阿糖胞苷能促进姜黄素诱导的细胞自噬,作用优于姜黄素单用组,可能与两药联合上调beclin-1和LC3-Ⅱ的表达有关。AIM: To observe whether autophagy occurs in curcumin-induced human acute myeloid leukemia KG1 a cells in the presence of chemotherapeutic drug cytarabine and the possible mechanism. METHODS: KG1 a cells were cultured in vitro. The ultrastructural changes of the cells were observed under transmission electron microscope. Autophagy was detected by acridine orange staining. The cell viability was measured by MTT assay. The cell cycle distribution was analyzed by flow cytometry. The expression of autophagy-related molecules beclin-1 and LC3 at mRNA and protein le-vels was determined by RT-qPCR and Western blot. RESULTS: Curcumin dose-dependently inhibited the viability of KG1 a cells(P<0.05). The growth inhibition rate in combination group was significantly higher than that in single reagent group and control group(P<0.01). Electron microscopical observation showed that curcumin induced the occurrence of autophagosomes, and cytarabine increased curcumin-induced autophagosomes. Acridine orange staining showed that the combined treatment with cytarabine increased the autophagy induced by curcumin, and the number of autophagic acid vesicles and cells containing autophagic acid vesicles were increased. Curcumin blocked the cell cycle in the G0/G1 phase. The mRNA expression levels of beclin-1 and LC3 in combination group were significantly higher than those in single reagent group and control group(P<0.01). The results of Western blot showed that the protein expression of beclin-1 was significantly up-regulated in combination group(P<0.05), and the ratio of LC3-II/LC3-I was higher than that in control group(P<0.01). CONCLUSION: Curcumin inhibits the viability of KG1 a cells and induces autophagy. Cytarabine promotes autophagy, which is superior to curcumin alone. It may be related to the up-regulation of beclin-1 and LC3-II by the two reagents.

关 键 词：姜黄素 自噬 阿糖胞苷 KG1a细胞 急性髓系白血病 

分 类 号：R733.71[医药卫生—肿瘤] R363.2[医药卫生—临床医学]