CIK细胞联合表柔比星对肝癌的体内外杀伤作用研究

The anti-tumor effects of CIK cells combined with Epirubicin against Hepatocellular carcinoma

目的研究CIK细胞联合表柔比星对肝癌细胞BEL-7402的体内外杀伤效应。方法取健康人外周血单个核细胞,经IFN-γ、IL-1、IL-2、抗CD3单抗诱导CIK细胞成熟,流式细胞仪检测培养第1、7、14d的CIK细胞表型。CellCountingKit-8试剂盒(CCK-8)检测CIK细胞、表柔比星及两者联合对肝癌细胞BEL-7402的体外杀伤效应。用肝癌细胞BEL-7402建立裸鼠皮下移植瘤模型,分为生理盐水对照组、CIK治疗组、表柔比星组、CIK联合表柔比星治疗组(联合组),观察它们对裸鼠移植瘤生长的抑制作用。结果CIK细胞经多种细胞因子诱导培养后,CD3+CD56+细胞比例明显上升,14d时CD3+CD56+细胞比例达(46.83±2.53)%,与第1天比差异有显著性(P<0.01),细胞量扩增了近1000倍。联合组对肝癌细胞的杀伤率明显高于各单独治疗组,差异有显著性(P<0.05)。体内实验表明,CIK细胞联合表柔比星可明显抑制裸鼠肝癌细胞种植瘤的生长且无明显副作用。结论细胞因子活化杀伤细胞疗法联合化疗能显著抑制体内外肝癌细胞BEL-7402的生长,为肝癌的生物化疗提供了实验室依据。

[ Objective ] To investigate the inhibitory effects of cytokine induced-kiUer （CIK） cells combined with Epirubicin on the expansion of Hepatocellular carcinoma cell line BEL-7402 in vitro and in vivo. [Methods] CIK cells were generated in vitro by stimulation of peripheral blood mononuclear cells subsets with interferon-gamma （IFN-γ）, IL-1, IL-2 and anti-CD3 monoclonal antibody. Phenotype analysis of CIK cells on the 1, 7, 14 days was performed with flow cytometer （FCM）. The specific cytotoxicities of CIK ceils combined with Epirubicin against Hepatocellular carcinoma cell line BEL-7402 was determined by Cell Counting Kit-8. Hepatocellalar carcinoma cell line BEL-7402 were injected subcutaneously into nude mice. On the 14th day, normal saline （control group）, CIK group, Epirubiein group and combination group were administered respectively. Tumor volume was measured after treatments and tumor inhibitory was calculated. [Results] Amount of CD3^＋CD56^＋ ceils were increased from （2.66 ± 0.42）% to （46.83 ± 2.53）% with elevated absolute amount over 1 000 times after 2 week＇s culture. Combined treatment of the cells resulted in significantly higher killing rate than that in the other groups （P 〈0.05）. Volume of implanted tumor in combination group was significantly smaller than in the other groups （P 〈0.05）. [ Conclusion ] CIK combined with Epirubicin can inhibit the growth of Hepatoeellular carcinoma ceil line BEL-7402 in vitro and in vivo.