Olaparib对HL-60细胞NKG2D配体表达的调节作用及相关机制研究  

Effects and Mechanism of PARP Inhibitor Olaparib on the Expression of NKG2D Ligands in HL-60 Cells

作　　者：朱志超[1] 白煜[1] 卢绪章[2] 戚春建[1] ZHU Zhi-Chao;BAI Yu;LU Xu-Zhang;QI Chun-Jian(Laboratory Center,The Affiliated Hospital of Nanjing Medical University,Changzhou 213000,Jiangsu Province,China;Department of Hematology,Changzhou No.2 People′s Hospital,The Affiliated Hospital of Nanjing Medical University,Changzhou 213000,Jiangsu Province,China)

机构地区：[1]南京医科大学附属常州市第二人民医院中心实验室,江苏常州213000 [2]南京医科大学附属常州市第二人民医院血液科,江苏常州213000

出　　处：《中国实验血液学杂志》2020年第6期1826-1830,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(81672799);常州市卫生计生委青年人才科技项目(QN201821);南京医科大学科技发展基金面上项目(2017NJMU044)。

摘　　要：目的:研究Olaparib对人急性髓系白血病细胞株HL-60细胞表面NKG2D配体表达的调节作用,并初步探索其内在调控机制。方法:对数生长期的HL-60细胞经不同浓度Olaparib(1.25、2.5、5、10μmol/L)作用24、48 h后,采用流式细胞术检测细胞表面NKG2D配体表达情况;Western blot检测HL-60细胞内ERK蛋白表达变化情况;CFSE/PI法检测NK细胞对HL-60细胞的杀伤作用。结果:10μmol/L Olaparib作用24和48 h均可上调HL-60细胞表面NKG2D配体的表达,5μmol/L Olaparib作用48 h可诱导ULBP-2、ULBP-3表达上调;Western blot检测结果显示,Olaparib处理后的HL-60细胞内ERK磷酸化水平增强。Olaparib可增强NK细胞对HL-60细胞的杀伤作用,但ERK抑制剂可下调NK细胞对HL-60细胞的杀伤作用。结论:Olaparib可上调HL-60细胞表面NKG2D配体表达,增强NK细胞对其的杀伤作用,其机制可能与Olaparib促进ERK磷酸化表达有关。Objective:To investigate the regulatory effects of Olaparib on natural killer cell activating receptor(NKG2D)ligands expression on human acute myeloid leukemia(AML)cell line HL-60,and to explore the molecular mechanism of Olaparib on HL-60 cells.Methods:After HL-60 cells in logarithmic growth phase were treated with Olaparib at different concentrations for different times(24,48 h),the expression of NKG2D ligand on the surface of HL-60 cells was detected by flow cytometry.Western blot was used to dectect the expression of ERK expression in HL-60 cells.The killing effect of NK cells to HL-60 cells was detected by CFSE/PI method.Results:10μmol/L Olaparib could upregulate the expression of NKG2D ligand on the surface of HL-60 cell at 24 and 48 hours,while 5μmol/L Olaparib could induce up-regulation of the expression of ULBP-2 and ULBP-3 at 48 hours.Western blot analysis showed that ERK phosphorylation of HL-60 cells was enhanced after treating with Olaparib.The killing effect of NK cells to HL-60 cells could be enhanced by Olaparib,however,ERK inhibitor could suppress the killing effect of NK cells to HL-60 cells.Conclusion:Olaparib can upregulate NKG2D ligands expression on the surface of HL-60 cells and enhance the cytotoxicity of NK cell to HL-60 cells.The mechanism may be related to Olaparib promoting ERK phosphorylation expression.

关 键 词：OLAPARIB PARP抑制剂 NKG2D配体 NK杀伤 急性髓系白血病 

分 类 号：R733.71[医药卫生—肿瘤]