辣椒素对人γδT细胞杀伤人骨肉瘤HOS细胞的影响

Effect of capsaicin on killing activity of γδT cells against human osteosarcoma HOS cells

探讨辣椒素对人γδT细胞体外增殖及杀伤骨肉瘤细胞的影响及作用机制。分离健康人PBMC，加入到含异戊烯焦磷酸（isopentenyl pyrophosphate，IPP）和IL-2的DMEM-F12完全培养基中诱导培养γδT细胞。不同浓度的辣椒素作用于γδT细胞48h后，CCK-8法检测各药物浓度组γδT细胞的增殖能力，采用流式细胞术检测各组γδT细胞穿孔素、颗粒酶B、CD107a及IFN-γ的表达，用LDH释放法检测各组γδT细胞对骨肉瘤HOS细胞的体外杀伤活性。结果显示，培养7d时各组帕T细胞纯度达到了（85．33±6．07）％。浓度为3．2～50μmol／L的辣椒素能显著促进帕T细胞的增殖，在浓度为25μmol／I。时其穿孔素、颗粒酶B、CD107a及IFN-γ的表达均显著高于对照组，且对人骨肉瘤HOS细胞的体外杀伤活性显著增强。以上结果提示辣椒素在体外能通过促进γδT细胞穿孔素、颗粒酶B和IFN-γ的表达上调显著增强其抗骨肉瘤细胞活性。

To study the effect of eapsaicin on proliferation and killing activity of γδT cells against osteosarcoma cells in vitro.γδT ceils derived from healthy donors were cultured in vitro in DMEM-F12 completed medium containing IPP and IL-2 for 7 d, and then co-cultured with different concentrations of eapsaicin for 48 h. Proliferation of each group γδTcells were detected by CCK-8 method. The phenotype was identified by flow cytometer. Expression of perforin, granzyme B, CD107a and IFN-γ produced by γδT cells were verified by flow cytometer. The cytotoxic activity of γδTcells against HOS cells were analyzed by LDH release assay. After being cultured for 7 d, the percentage of γδT cells reached （85.33±6.07） %. Compared with the control group that has not been treated with capsaicin, the proliferation of γδT cells incubated with 3.2- 50μmol/L capsaicin for 48 h were enhanced significantly. Furthermore, killing activity against HOS cells and expressions of perforin, granzyme B, CD107a and IFN-γ by γδT cells incubated with 25μmol/L eapsaicin were markedly higher than the control group. The results suggest that eapsaiein could significantly increase γδT cells eyotoxic activity against osteosarcoma HOS ceils, and up-regula- tion of perforin, granzyme B and IFN-γ expression may be the important molecular mechanism for killing of osteosarcoma cells.