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目的研究甘露聚糖结合凝集素(MBL)对白血病细胞系U937凋亡的影响。方法不同浓度MBL处理U937细胞后,应用CCK-8法分析细胞增殖情况,Hoechst33258染色观察细胞核及染色质,AnnexinV/PI双染流式细胞术分析细胞凋亡率,real-timePCR和免疫印迹分析凋亡相关基因及蛋白表达。结果 30～50μg/mlMBL培养72h,U937细胞增殖明显受抑,出现不同程度核固缩、核碎裂;随MBL浓度升高或作用时间延长,凋亡细胞数逐渐增多;Fas、Caspase-3mRNA、Fas和FasL蛋白表达量升高,Caspase-3和多腺苷二磷酸多聚酶(PARP)蛋白被剪切激活或失活。结论 MBL可诱导白血病细胞U937细胞凋亡,其机制与上调Fas表达、剪切Caspase-3和PAPR有关。
Objective To study the effect of mannan-binding lectin (MBL) on the apoptosis of leukemia cell line U937. Methods The proliferation of U937 cells treated with different concentrations of MBL was determined by CCK-8 assay. The cell nuclear and chromatin were stained with Hoechst33258 and the apoptosis rate was analyzed by flow cytometry with Annexin V / PI double staining. Real-time PCR and Western blot analysis Related genes and protein expression. Results The proliferation of U937 cells was significantly inhibited by 30 ~ 50μg / ml MBL for 72h, and nuclear pyknosis and nuclear fragmentation appeared in different degrees. The number of apoptotic cells gradually increased with the increase of MBL concentration or the prolonged action time. The expressions of Fas, Caspase-3 mRNA, Fas and FasL protein expression increased, Caspase-3 and poly-adenosine diphosphate polymerase (PARP) protein is sheared activation or inactivation. Conclusion MBL can induce the apoptosis of leukemia cell line U937 by up-regulating the expression of Fas, cleaving Caspase-3 and PAPR.