沉默CSE1L对急性髓系白血病细胞阿糖胞苷敏感性的影响及作用机制研究  

Effect of CSE1L silencing on cytarabine sensitivity of acute myeloid leukemia cells

作　　者：刘小玉 李旭东 关坤萍[3] LIU Xiaoyu;LI Xudong;GUAN Kunping(Department of Biochemistry and Molecular Biology,School of Basic Medicine,Shanxi Medical University,Taiyuan 030001,China;The Second Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Laboratory,The Second Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学基础医学院生物化学与分子生物学教研室,山西太原030001 [2]山西医科大学第二临床医学院,山西太原030001 [3]山西医科大学第二医院检验科,山西太原030001

出　　处：《中国病理生理杂志》2023年第2期269-275,共7页Chinese Journal of Pathophysiology

基　　金：山西省重点研发计划项目(No.201903D321093)。

摘　　要：目的:探讨敲减染色体分离1样蛋白(CSE1L)对急性髓系白血病细胞株THP-1细胞生物学行为及阿糖胞苷(Ara-C)敏感性的影响,并分析相关作用机制。方法:采用RT-qPCR法检测CSE1L的mRNA表达水平,流式细胞术检测细胞周期及凋亡水平,LinkedOmics及DAVID数据库进行基因本体论(gene ontology,GO)功能注释和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析,Western blot法检测细胞周期素D1(cyclin D1)、原癌基因c-Myc、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的相对表达水平。结果:与对照组相比,空载(sh-NC)组及敲减(sh-CSE1L)组标记绿色荧光蛋白(GFP)的THP-1细胞占比均>90%。与空载组相比,敲减组CSE1L的mRNA相对表达量降低至40%,凋亡率显著升高(P<0.01)。敲减组比空载组G0/G1期细胞占比升高(P<0.05),S期细胞占比降低(P<0.05),而G2/M期无明显差异。在1μmol/L阿糖胞苷处理24 h后,敲减组细胞活力最低(P<0.05),凋亡率最高(P<0.01)。此外,敲减CSE1L可下调Bcl-2、cyclin D1和c-Myc蛋白表达水平,上调Bax蛋白表达水平(P<0.05)。生物信息学分析显示CSE1L主要参与细胞周期等通路。结论:敲减CSE1L可阻滞THP-1细胞周期,诱导细胞凋亡,提高细胞对阿糖胞苷的敏感性,其可能通过调控细胞周期相关分子发挥作用。AIM:To investigate the effect of chromosome segregation 1-like (CSE1L) on biological behavior and cytarabine (Ara-C) sensitivity of acute myeloid leukemia THP-1 cells,and to analyze its mechanism.METHODS:The THP-1 cells were divided into control group,negavtive control (sh-NC) group,sh-CSE1L group,sh-NC+Ara-C group,and sh-CSE1L+Ara-C group.The mRNA expression of CSE1L was detected by RT-qPCR.Cell cycle and apoptosis were detected by flow cytometry.Gene ontology (GO) functional annotation and KEGG pathway enrichment analysis were performed by LinkedOmics and DAVID databases.The protein levels of cyclin D1,c-Myc,Bax and Bcl-2 were detected by Western blot.RESULTS:The expression of green fluorescent protein (GFP) was more than 90%in both the sh-NC and sh-CSE1L groups when compared with control group.Compared with sh-NC group,the mRNA expression level of CSE1L was reduced to 40%in sh-CSE1L group,and the apoptosis rate was significantly higher (P<0.01).The cell cycle G0/G1phase was significantly higher (P<0.05),and the S phase was significantly lower (P<0.05) in sh-CSE1L group than in sh-NC group,while there was no significant difference in G2/M phase.Cell viability was the lowest (P<0.05) and apoptosis was the highest (P<0.01) in sh-CSE1L group after treated with Ara-C.In addition,silencing of CSE1L downregulated Bcl-2,cyclin D1 and c-Myc protein levels (P<0.05),but up-regulated Bax protein level (P<0.05).Bioinfor?matic analysis concluded that CSE1L is mainly involved in the cell cycle and other pathways.CONCLUSION:Knock?down of CSE1L blocks THP-1 cell cycle,induces apoptosis,and improves the sensitivity to Ara-C,which may act by regu?lating cell cycle-related molecules.

关 键 词：阿糖胞苷 急性髓系白血病 染色体分离1样蛋白 细胞周期 细胞凋亡 

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