沙利度胺对白血病细胞NKG2D配体表达及NK细胞杀伤敏感性的影响  

Effects of thalidomide on expression of NKG2D ligands in leukemia cells and cytotoxicity sensitivity of NK cells

作　　者：邓黎黎 张岩 张壮苗 梅瑜 颜榕辛 罗伟 李洪芳 DENG Lili;ZHANG Yan;ZHANG Zhuangmiao;MEI Yu;YAN Rongxin;LUO Wei;LI Hongfang(Department of Hematology and Oncology,Sanya People's Hospital,Sanya Hainan 572000,China)

机构地区：[1]三亚市人民医院血液肿瘤内科,海南三亚572000

出　　处：《临床与病理杂志》2022年第1期9-14,共6页Journal of Clinical and Pathological Research

基　　金：三亚市医疗卫生科技创新项目(2014YW09)。

摘　　要：目的:探究沙利度胺对白血病细胞自然杀伤细胞及活性受体D(natural killer cell group 2 member D,NKG2D)配体表达及自然杀伤细胞(natural killer,NK)杀伤敏感性的影响。方法:取对数生长期白血病细胞株HL-60、K562细胞,以不同浓度沙利度胺作用48h,并设置未经沙利度胺处理的细胞为对照组,采用细胞计数实验(cell counting kit-8,CCK-8)检测沙利度胺对细胞的半抑制浓度(half-inhibitory concentration,IC50),实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-PCR)和流式细胞术检测细胞NKG2D配体[MHC-I类链相关分子A(major histocompatibility complex classⅠchain-related protein A,MICA)、MICB]及人巨细胞病毒UL16蛋白的结合蛋白[(UL16-binding proteins,ULBPs:ULBP1、ULBP2、ULBP3)]表达情况,乳酸脱氢酶(lactate dehydrogenase,LDH)释放法检测沙利度胺对NK-92MI细胞的杀伤效率。结果:沙利度胺干预HL-60、K562细胞,IC50分别为30.06、31.95μg/mL,且随着沙利度胺浓度的升高,抑制作用越明显;与对照组比较,沙利度胺组MICB、ULBP1、ULBP2基因mRNA水平明显升高(P<0.05),但MICA和ULBP3 mRNA水平无明显变化(P>0.05);与对照组比较,沙利度胺组MICB、ULBP1、ULBP2表达明显升高(P<0.05),但MICA和ULBP3表达无明显变化(P>0.05);效靶比分别为1:1、5:1、10:1时,与对照组比较,沙利度胺组NK-92MI对细胞的杀伤效率明显升高(P<0.05)。结论:沙利度胺可能通过提高NKG2D配体MICB、ULBP1、ULBP2表达,提高HL-60、K562细胞对NK-92MI的杀伤敏感性。Objective:To explore the effects of thalidomide on the expression of natural-killer group 2 member D(NKG2D)ligands in leukemia cells and cytotoxicity sensitivity of natural killer cell(NK).Methods:The leukemia cell lines HL-60 and K562 cells in logarithmic growth phase were collected and treated with different concentrations of thalidomide for 48 h.The cells without thalidomide treatment were collected as control group.The half-inhibitory concentration(IC50)of thalidomide on cells was detected by cell counting kit-8(CCK-8).The expressions of NKG2D ligands[major histocompatibility complex class I chain-related protein A(MICA),MICB],UL16-binding proteins(ULBPs:ULBP1,ULBP2,ULBP3)were detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR)and flow cytometry.The cytotoxicity efficiency of thalidomide on NK-92MI cells was detected by lactate dehydrogenase(LDH)release method.Results:IC50 values for HL-60 and K562 cells were 30.06 and 31.95μg/mL,respectively.With the increase of thalidomide concentration,the inhibitory effects were more significant.Compared with the control group,mRNA levels of MICB,ULBP1 and ULBP2 genes were significantly increased in the thalidomide group(P<0.05),but there was no significant change in mRNA levels of MICA and ULBP3(P>0.05).Compared with the control group,expressions of MICB,ULBP1 and ULBP2 were significantly increased in the thalidomide group(P<0.05),but there was no significant change in the expression of MICA and ULBP3(P>0.05).Themultiplicity of infection values were 1:1,5:1 and 10:1,respectively.Compared with the control group,cytotoxicity efficiency of NK-92MI on cells was significantly increased in thalidomide group(P<0.05).Conclusion:Thalidomide may improve the cytotoxicity sensitivity of HL-60 and K562 cells to NK-92MI by increasing the expressions of NKG2D ligands(MICB,ULBP1,ULBP2).

关 键 词：沙利度胺 急性白血病 自然杀伤细胞 NKG2D配体 杀伤敏感性 

分 类 号：R733.7[医药卫生—肿瘤]