IL-27联合IL-15通过磷酸化STATs途径调控NK92细胞的抗肿瘤作用  

IL-27 in combination with IL-15 regulates anti-tumor effect of NK92 cells by phosphorylating the STATs pathway

作　　者：蒋亚楠 孙雨飞[1] 王坤[2] 付强[1,2,3] JIANG Yanan;SUN Yufei;WANG Kun;FU Qiang(Department of Immunology,Binzhou Medical University,Yantai 264003,Shandong,China;Department of Biological Pharmacy,Binzhou Medical University,Yantai 264003,Shandong,China;Shandong Cellogene Pharmaceutics Co.,LTD.,Yantai 264003,Shandong,China)

机构地区：[1]滨州医学院免疫学教研室,山东烟台264003 [2]滨州医学院生物制药教研室,山东烟台264003 [3]山东思乐基医药科技有限公司,山东烟台264003

出　　处：《中国肿瘤生物治疗杂志》2021年第3期261-268,共8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);烟台市高端人才引进“双百计划”专项经费资助。

摘　　要：目的:探讨IL-27联合IL-15对NK92细胞抗肿瘤作用的影响及其分子和信号通路机制。方法:将高表达IL-15的NK92(IL-15-NK92)细胞分别置于不同质量浓度的IL-27(0、10、20、30及60 ng/ml)下培养24 h,ELISA法检测IL-27对IL-15-NK92细胞分泌IL-15的影响,Transwell法检测IL-27对IL-15-NK92细胞迁移作用的影响,CCK-8法检测IL-27对IL-15-NK92细胞增殖能力的影响,流式细胞术检测IL-15-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,LDH法检测IL-15-NK92细胞对多种血液肿瘤和实体瘤细胞的杀伤作用,WB检测STATs通路相关蛋白的表达及其磷酸化水平。结果:30 ng/ml的IL-27可以促进IL-15-NK92细胞IL-15的分泌(P<0.01)、明显增强其迁移能力(P<0.05),但明显抑制其增殖能力(P<0.05);并且显著促进IL-15-NK92细胞表面受体NKG2D、NKp30和NKp46的表达(均P<0.05)、明显促进穿孔素分泌(P<0.05)但不影响颗粒酶B的分泌(P>0.05)。30 ng/ml的IL-27可显著促进IL-15-NK92细胞对多种血液肿瘤和实体瘤细胞的杀伤能力(P<0.05或P<0.01),并且上调STAT1、STAT3、及STAT5蛋白的磷酸化水平(均P<0.01)。结论:IL-27可增强IL-15-NK92细胞对实体肿瘤细胞和血液肿瘤细胞的杀伤作用,该作用与IL-27上调IL-15-NK92细胞中JAK-STAT通路相关蛋白STAT1、STAT3、STAT5磷酸化水平和促进该细胞中多种活化性受体相关。Objective: To investigate the effect of IL-27 in combination with IL-15 on the anti-tumor effects of NK92 cells and the possible molecular and signaling mechanisms. Methods: NK92 cells with high IL-15 expression(IL-15-NK92 cells) were cultured in different mass concentrations of IL-27(0, 10, 20, 30 and 60 ng/ml) for 24 h. The effects of IL-27 on IL-15 secretion, migration and proliferation of IL-15-NK92 cells were detected by ELISA, Transwell and CCK-8 assay, respectively. Flow cytometry was used to detect the expression levels of IL-15-NK92 cell surface receptors NKG2 D, NKp30 and NKp46, as well as the secretion levels of perforin and granzyme B. LDH method was used to detect the cytotoxic effect of IL-15-NK92 cells on hematologic tumor cells and solid tumor cells, and WB was used to detect the expressions and phosphorylation level of STATs pathway-related proteins. Results: IL-27 at the concentration of 30 ng/ml promoted IL-15-NK92 cells secreting IL-15(P<0.01), significantly enhanced the cell migration(P<0.05) but inhibited the proliferation of IL-15-NK92 cells(P<0.05). 30 ng/ml IL-27 could significantly promote the expressions of NKG2 D, NKp30 and NKp46 on surface of IL-15-NK 92 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 significantly enhanced the cytotoxicity of IL-15-NK92 cells against hematologic malignancies and solid tumor cells(P<0.05 or P<0.01), and up-regulated the phosphorylation levels of STAT1, STAT3 and STAT5(all P<0.01). Conclusion: IL-27 can enhance the cytotoxicity of IL-15-NK92 cells against hematologic tumor cells and solid tumor cells, which might be related with its upregulation of phosphorylation level of STAT1, STAT3 and STAT5 in JAK-STAT pathway and multiple activating receptors in IL-15-NK92 cells.

关 键 词：IL-27 IL-15 NK92细胞 抗肿瘤 JAK-STAT通路 

分 类 号：R7350.51[医药卫生—肿瘤] R392.1[医药卫生—临床医学]