Nucleostemin下调联合雷帕霉素对HL-60细胞自噬与凋亡影响的初步研究  

Preliminary Study on the Effect of Silencing Nucleostemin Combined with Rapamycin on Autophagy and Apoptosis of HL-60 Cells

作　　者：王亚奇 高晓娟 岳保红 WANG Ya-Qi;GAO Xiao-Juan;YUE Bao-Hong(Department of Laboratory Medicine,The First Affliated Hospital of Zhengzhou University,Key Clinical Laboratory of Henan Province,Faculty of Laboratory Medicine,Zhengzhou University;Department of Laboratory Medicine,The Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Prorince,China)

机构地区：[1]郑州大学第一附属医院检验科,河南省检验医学重点实验室,郑州大学医学检验系 [2]郑州大学第五附属医院检验科,河南郑州450052

出　　处：《中国实验血液学杂志》2023年第6期1629-1634,共6页Journal of Experimental Hematology

摘　　要：目的:研究Nucleostemin(NS)下调联合雷帕霉素对HL-60细胞自噬和凋亡的影响并探讨其在HL-60细胞中的作用。方法:通过NS-RNAi-GV248重组慢病毒载体转染HL-60细胞后检测NS蛋白的表达。采用流式细胞术检测沉默NS和/或雷帕霉素处理24、48 h后HL-60细胞的凋亡变化;采用Western blot技术检测各组细胞中NS、LC3、p62、BCL-2、Bax蛋白的表达。结果:重组慢病毒载体成功下调HL-60细胞NS表达。经雷帕霉素处理后,细胞凋亡率均明显增加(P<0.05),且48 h凋亡更明显。与NS敲低组、雷帕霉素组相比,NS下调联合雷帕霉素组处理48 h后,其凋亡明显增加(P<0.05),LC3-II/LC3-I比值显著增加(P<0.05),p62蛋白表达下降更显著(P<0.05),并且BCL-2/Bax比值减低更明显(P<0.05)。结论:NS下调联合雷帕霉素可增强HL-60细胞的凋亡和自噬,并且其诱导HL-60细胞的凋亡可能与BCL-2、Bax蛋白的表达有关。Objective:To investigate the effects of knocking down nucleostemin(NS)combined with rapamycin(RAPA)on autophagy and apoptosis in HL-60cells,and to explore its role in HL-60cells.Methods:The expression of NS protein was detected using Western blot,after transfection of HL-60cells was achieved by the recombinant lentviral vector NS-RNAi-GV248.Flow cytometry was used to detect changes in cells apoptosis after NS silencing/rapamycin for24,48hours,and the expressions of NS,LC3,p62,BCL-2and Bax proteins in cells were detected by Western blot.Results:The expression of NS in HL-60cells was successfully down-regulated by recombinant lentiviral vector.After treatment with rapamycin for24and48h,the apoptosis rate of cells in each group increased(P<0.05),and the apoptosis was more obvious at48hours.Compared with the NS silencing group or rapamycin group,after treated with NS down-regulation combined with rapamycin for48hours,the apoptosis of HL-60cells was significantly increased(P<0.05),LC3-II/LC3-I ratio was significantly increased(P<0.05),p62protein expression was significantly decreased(P<0.05),and BCL-2/Bax ratio was significantly decreased(P<0.05).Conclusion:NS down-regulation combined with rapamycin can enhance the apoptosis and autophagy of HL-60cells,and the induction of apoptosis of HL-60cells may be related to the expression of BCL-2and Bax proteins.

关 键 词：核干细胞因子 HL-60细胞 雷帕霉素 自噬 凋亡 

分 类 号：R733.71[医药卫生—肿瘤]