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
| 作者: |
1邓黎黎,
1张岩,
1张壮苗,
1梅瑜,
1颜榕辛,
1罗伟,
1李洪芳
1 三亚市人民医院血液肿瘤内科，海南 三亚 572000 |
| 通讯: |
邓黎黎
Email: dengll1984108@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2022.01.002 |
| 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48 h，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 class I chain-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、31.95 μg/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。
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
Effects of thalidomide on expression of NKG2D ligands in leukemia cells and cytotoxicity sensitivity of NK cells
CorrespondingAuthor: DENG Lili Email: dengll1984108@163.com
DOI: 10.3978/j.issn.2095-6959.2022.01.002
Foundation: This work was supported by the Medical and Health Science and Technology Innovation Project of Sanya, China (2014YW09).
Abstract
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 31.95 μg/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).
Keywords：
thalidomide; acute leukemia; natural killer cell; NKG2D ligand; cytotoxicity sensitivity