【摘要】 目的:研究PD-1阻断增强体外扩增的NK细胞对人非小细胞肺癌(NSCLC)杀伤作用的研究。方法:采用来自健康人群的外周静脉血的单个核细胞以及NSCLC细胞(HCC827细胞)进行体外扩增,随后PD-1进行阻断增强,使用培养10 d的PD-1(0.25、0.5、1μmol/L)阻断增强NK细胞以及未进行PD-1阻断增强NK细胞,分别培养24 h、48 h、96 h,分析PD-1阻断增强NK细胞的细胞毒性、PD-1阻断增强NK细胞颗粒酶B、穿孔素、CD107a表达水平以及不同浓度PD-1阻断增强NK细胞对HCC827细胞的抑制率之间的差异。结果:通过对离心管1～3中的PD-L1、PD-L2表达水平以及对NK细胞的PD-1阻断增强作用进行比较,其PD-L1及PD-L2表达水平显著下降;不同浓度PD-1阻断增强NK细胞颗粒酶B、穿孔素、CD107a表达水平之间的差异存在统计学意义(P<0.05),在0.5μmol/L PD-1阻断增强NK细胞的细胞颗粒酶B、穿孔素、CD107a表达水平达到高峰。0.5μmol/L PD-1阻断增强NK细胞对HCC827细胞的抑制率显著高于其他两组。结论:0.5μmol/L PD-1阻断增强NK细胞的阻断作用其中人NSCLC的细胞阻断作用显著,其对肿瘤细胞的杀灭作用的机制与活化型受体的升高有关。更多还原

【Abstract】 Objective:To study the killing effect of PD-1 on human non-small cell lung cancer(NSCLC).Methods:Peripheral blood mononuclear cells and NSCLC cells(HCC827 cells)from the peripheral venous blood of healthy people were used for in vitro amplification,followed by PD-1 blockade and enhancement.The above NK cells were expanded in vitro,and then PD-1 was blocked and enhanced. PD-1 blocked and enhanced NK cells were cultured for 10 d with concentration of 0.25,0.5 and 1 μmol/L,respectively,for 24 h,48 h and 96 h. PD-1 blocked and enhanced the cytotoxicity of NK cells,PD-1 blocked and enhanced the expression level of granzyme B,perforin and CD107 a of NK cells PD-1 blockade enhanced the difference of inhibition rate between NK cells and hcc827 cells.Results:By comparing the expression levels of PD-L1 and PD-L2 in centrifuge tube 1～3,the expression levels of PD-L1 and PDL2 in NK cells decreased significantly by blocking and enhancing the PD-1;the difference of PD-1 in different concentrations enhanced the expression levels of granzyme B,perforin and CD107 a in NK cells was statistically significant(P<0.05). The expression levels of granzyme B,perforin and CD107 a reached the peak. 0.5 μmol/L PD-1 block enhanced the inhibition rate of NK cells on hcc827 cells significantly higher than the other two groups.Conclusion:0.5 μmol/L PD-1 can enhance the blocking effect of NK cells on NSCLC cells,and the mechanism of its killing effect on tumor cells is mainly related to the increase of activated receptor.更多还原