IL-27通过促进活化性受体表达和STATs磷酸化增强NK92细胞的抗肿瘤作用  

IL-27 promotes anti-tumor effect of NK92 cells by up-regulating expression of activated receptors and STATs phosphorylation

作　　者：孙雨飞[1] 蒋亚楠 于淮海 王玉芳 付强[1,3] SUN Yufei;JIANG Yanan;YU Huaihai;WANG Yufang;FU Qiang(Department of Immunology,Binzhou Medical University,Yantai 264003,Shandong,China;Department of Clinical Laboratory,Fushan People's Hospital,Yantai 265500,Shandong,China;University of North Carolina Lineberger Cancer Center,Chapel Hill 27514,USA)

机构地区：[1]滨州医学院免疫学教研室,山东烟台264003 [2]烟台市福山区人民医院检验科,山东烟台265500 [3]北卡罗莱纳大学Lineberger肿瘤中心,美国教堂山27514

出　　处：《中国肿瘤生物治疗杂志》2019年第5期500-505,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81370730;No.81571512);山东省自然科学基金重点项目资助(No.ZR2015JL027)~~

摘　　要：目的:探讨IL-27影响NK92细胞抗肿瘤作用的分子和信号通路的机制。方法:将NK92细胞分别置于不同质量浓度的IL-27(10、20、30及60 ng/ml)下培养24 h,LDH法检测NK92细胞对多种血液肿瘤和实体瘤细胞的杀伤作用,流式细胞术检测NK92细胞表面受体NKG2D、NKp30和NKp46的表达水平以及穿孔素和颗粒酶B的分泌水平,WB检测STATs蛋白的表达和磷酸化水平。建立重度免疫缺陷型小鼠的前列腺癌(DU145细胞)模型,使用IL-27和NK92细胞联合局部治疗,评估其抗肿瘤活性。结果:10、20及30 ng/ml浓度的IL-27均可显著促进NK92细胞对靶细胞的杀伤能力,且30 ng/ml的IL-27对其细胞毒的促进作用最强(P<0.05或P<0.01)。30 ng/ml IL-27能显著促进NK92细胞表面受体NKG2D、NKp30和NKp46的表达(均P<0.05)、明显促进NK92分泌穿孔素(P<0.05),但不影响颗粒酶B的分泌(P>0.05);上调STAT1、STAT3及STAT5蛋白的磷酸化水平(均P<0.01)。IL-27和NK92细胞局部联合治疗可明显延长前列腺癌荷瘤小鼠生存期(P<0.05)。结论:IL-27可增强NK92细胞对实体肿瘤细胞和血液肿瘤细胞的杀伤作用,两者联合治疗可明显延长荷瘤小鼠生长期,该作用与IL-27上调NK92细胞中JAK-STAT通路STAT1、STAT3、STAT5磷酸化水平和促进细胞中多种活化性受体相关。Objective:To investigate the molecular and signal pathway mechanism of Interleukin-27 affecting the anti-tumor effect of NK92 cells.Methods:NK92 cells were cultured with different concentrations of IL-27(10,20,30 and 60 ng/ml)for 24 hours.The cytotoxicity of NK92 cells to target cells was detected by LDH assay.The expressions of NKG2 D,NKp30 and NKp46 on the surface of NK92 cells and the secretion of perforin and granzyme B were detected by Flow cytometry.The expression and phosphorylation level of STATs protein was detected by WB.The DU145 cell transplanted tumor model of prostatic carcinoma in NOD-PrkdcscidIl2 rgem1/Smoc mice was established and treated with the combination of NK92 cells and IL-27 to evaluate their anti-tumor efficacy.Results:IL-27 at concentrations of 10,20 and 30 ng/ml could significantly increase the cytotoxicity of NK92 cells to target cells,and 30 ng/ml exerted the best effect(P<0.05 or P<0.01).30 ng/ml IL-27 could significantly promote the expressions of NKG2 D,NKp30 and NKp46 on surface of NK92 cells,as well as elevate the secretion of perforin(all P<0.05),but didn’t affect the secretion of granzyme B(P>0.05);moreover,it also up-regulated the phosphorylation of STAT1,STAT3 and STAT5 protein(all P<0.01).The combined treatment of IL-27 and NK92 cells obviously extended the survival time of tumor-bearing mice(P<0.05).Conclusions:IL-27 can promote the cytotoxicity of NK92 cells against solid tumor cells and blood tumor cells by promoting expressions of NKG2 D,NKp30 and NKp46 on the surface of NK92 cells and the secretion of perforin,which might be related with the phosphorylation of STAT1,STAT3 and STAT5 in JAKSTAT pathway.

关 键 词：IL-27 NK92 JAK-STATs 抗肿瘤 

分 类 号：R730.3[医药卫生—肿瘤] R392.12[医药卫生—临床医学]