DC与CIK共培养对骨肉瘤细胞的杀伤作用

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

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。

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.