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 NKG2D, 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 NKG2D, 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.

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