目的：观察TIM-3 在NK/T细胞淋巴瘤（natural killer/T cell lymphoma, NK/TCL）细胞株中的表达及其配体galectin-9（GAL-9）诱导瘤细胞凋亡的作用及其部分机制。方法：Western blotting 测定NK/TCL细胞株SNK-1、SNK-6、SNT-8 和健康人外周血NK细胞中TIM-3 的表达；采用不同质量浓度重组人GAL-9（rhGAL-9）作用于NK/TCL细胞株24 h 后，通过CCK-8 法检测细胞增殖活性；流式细胞术Annexin-V/PI 双染法检测瘤细胞凋亡；Western blotting 检测caspase-3、PARP及其剪接体和MAPK信号通路蛋白磷酸化表达水平变化。结果：NK/TCL细胞株SNK-1、SNK-6 和SNT-8 的TIM-3 表达较健康人外周血NK细胞显著增高，其中SNK-1、SNK-6 细胞株与对照组相比差异具有统计学意义（P<0.05）；CCK-8 检测结果显示，rhGAL-9 对NK/TCL细胞增殖活力具有明显抑制作用，并有浓度依赖性；流式细胞术检测显示，rhGAL-9 可诱导3 种NK/TCL细胞株凋亡；Western blotting 结果显示，cleaved-caspase-3、cleaved-PARP蛋白表达量增高，MAPK通路中JNK蛋白磷酸化水平升高。结论：TIM-3 在NK/TCL细胞株中异常高表达，其配体GAL-9可诱导细胞凋亡，其机制可能与JNK磷酸化水平升高相关。

Objective: To identify the expression pattern of TIM-3 in natural killer/T-cell lymphoma (NK/TCL) cell lines, and to investigate the effect and mechanism of its ligand galectin-9 (GAL-9) inducing apoptosis of NK/TCL cell lines. Methods: Expression of TIM-3 in NK cell of peripheral blood from healthy donors and NK/TCL cell lines (SNK-1、SNK-6、SNT-8) was detected by Western blotting. After being treated with rhGAL-9 at various concentrations for 24h, the cell proliferation ability was analyzed with CCK-8 assay.Apoptosis ratio of the cells was determined by flow cytometry. Expressions of caspase-3, PARP and their cleavages were detected by Western blotting; moreover, phosphorylation levels of proteins in MAPK signaling pathway were also detected by Western blotting.Results: The expression of TIM-3 in SNK-1, SNK-6 and SNT-8 cell lines was significantly higher than that of NK cells from healthy donors (P<0.05). CCK-8 result showed that rhGAL-9 obviously inhibited the proliferation of NK/TCL cell lines in a concentration dependent manner. Flow cytometry showed that rhGAL-9 induced the apoptosis of NK/TCL cells; andWestern blotting proved that the expression of cleaved caspase-3, cleaved-PARP, and p-JNK in MAPK signaling pathway were significantly elevated. Conclusion: TIM-3 was over-expressed in NK/TCL cell lines, and its ligand galectin-9 induced cell apoptosis probably through the activation of JNK kinase pathways.

上海市教育委员会高原高峰学科建设计划（20152219）