Toll样受体7激动剂对K562细胞抑制作用的研究

Inhibitory Effect of Toll-like Receptor 7 Agonist on K562 Cells

本研究旨在探讨Toll样受体(TLR)7激动剂gardiquimod对人慢性髓系白血病细胞K562的抑制作用。以异戊烯焦磷酸法扩增人外周血γδT细胞。用不同浓度的gardiquimod处理γδT细胞和K562细胞48 h,用MTT法检测细胞增殖情况。以流式细胞术检测gardiquimod处理前后K562细胞内TLR7表达、细胞周期及凋亡的变化。用CCK-8试剂盒检测γδT细胞对K562的杀伤活性。结果表明,gardiquimod 0.69-11.0μg/ml可明显促进γδT细胞增殖,在5.5-11.0μg/ml浓度下抑制K562细胞增殖;gardiquimod处理K562细胞后未检测到凋亡,但细胞周期有明显变化,而且处理后的K562细胞更易被γδT细胞杀伤。结论:gardiquimod能够抑制K562细胞增殖,阻滞细胞周期,并增强其对γδT细胞杀伤的敏感性。

The aim of this study was to investigate the inhibitory effect of Toll-like receptor 7（TLR7） agonist gardiquimod on K562 cells.Human γδT cells from peripheral blood cells were amplified by isopentenyl pyrophosphate.The proliferation capacity of γδT cells and K562 cells were measured with MTT assay after treatment with different concentrations of gardiquimod.Cytotoxicity of γδT cells on K562 cells was detected by CCK-8 kit,and the intracellular expression of TLR7,cell cycle and apoptosis of K562 cells before and after treatment with gardiquimod were measured by flow cytometry.The results demonstrated that gardiquimod could significantly stimulate the proliferation of γδT cells,and inhibit proliferation of K562 cells under the concentration of 11.0 μg/ml for 48 h.The expression of TLR7 increased after treatment with gardiquimod.No apoptosis was observed,but there were significant changes in cell cycle,moreover the K562 cells treated with gardiquimod were more killed by γδT cells.It is concluded that the gardiquimod can inhibit the proliferation of K562 cells and enhance their sensitivity to killing activity of human γδT cells.