DC与CIK共培养对骨肉瘤细胞的杀伤作用

目的探讨树突状细胞(dendritic cells,DC)与细胞因子诱导的杀伤细胞(cytokine-induced killer cells,CIK)共培养对人骨肉瘤细胞的杀伤效应。方法从健康志愿者中分离外周血单个核细胞,制备DC和CIK,并以5∶1比例混合培养。采用CCK-8法和流式细胞术检测DC-CIK共培养对人骨肉瘤HOS细胞增殖和凋亡的影响;构建裸鼠移植瘤模型,观察DC-CIK共培养对裸鼠体内移植瘤生长的影响。结果流式细胞术检测结果显示,分化成熟的DC中CD83+和CD86+阳性细胞比例均高于未成熟的DC(P=0.009,0.006);DC-CIK共培养后,CD3+CD8+和CD3+CD56+阳性细胞比例均高于CIK单独培养组(P=0.004,0.004)。与DC-CIK共培养后,HOS细胞增殖能力受抑制,细胞凋亡能力增强(P<0.05);PD-L1的表达水平明显下调(P=0.006)。裸鼠移植瘤模型中,DC-CIK共培养可抑制HOS细胞诱导的移植瘤生长(P=0.008)。结论DC-CIK共培养能抑制人骨肉瘤HOS细胞增殖并诱导细胞凋亡,其作用机制可能与靶向调控PD-1/PD-L1信号通路有关。

Killing effect of DC-CIK co-culture on osteosarcoma cells

Objective To investigate the killing effect of dendritic cells (DC) co-cultured with cytokine induced killer (CIK) cells on human osteosarcoma cells. Methods The peripheral blood mononuclear cells were isolated from healthy volunteers, the DC and CIK were prepared and co-cultured at a ratio of 5∶1. CCK-8 assay and flow cytometry were used to detect the effects of DC-CIK on the proliferation and apoptosis of human osteosarcoma HOS cells. The model of transplanted tumor in nude mice was established to observe the effect of DC-CIK on the growth of transplanted tumor. Results The results of flow cytometry showed that the percentage of CD83+ and CD86+ positive cells in mature DC was higher than that in immature DC (P=0.009, 0.006). The percentage of CD3+CD8+ and CD3+CD56+ positive cells in DC-CIK co-culture group was higher than that in CIK group (P=0.004, 0.004). After co-cultured with DC-CIK, the proliferation ability of HOS cells was inhibited and the apoptosis ability was enhanced (P<0.05). The expression of PD-L1 was significantly down-regulated (P=0.006). In nude mice model, DC-CIK co-culture could inhibit the growth of HOS cell-induced transplanted tumor (P=0.008). Conclusions DC-CIK co-culture can inhibit the proliferation and induce apoptosis of human osteosarcoma HOS cells, and its mechanism may be related to the targeted regulation of PD-1/PD-L1 signaling pathway.