DC-GPC3联合CIK 细胞的体外抗肝癌作用

摘要： 目的 探讨磷脂酰肌醇蛋白聚糖-3 （GPC3） 基因转染的树突状细胞 （DC-GPC3） 与细胞因子诱导杀伤细胞（CIK） 共培养 （DCIK-GPC3） 后， 对 CIK 的生物活性及体外抗肝癌细胞作用。方法 流式细胞术检测 DCIK-GPC3、 DC-CIK 及 CIK 各组效应细胞免疫表型， MTT 法检测各组效应细胞培养上清液中白细胞介素 （IL） -2 生物活性， 酶联免疫吸附试验 （ELISA） 检测细胞培养上清液中 IL-2、 干扰素 γ （IFN-γ） 浓度。乳酸脱氢酶释放实验分别检测各组效应细胞对肝癌 HepG2 细胞的细胞毒活性。结果 DCIK-GPC3 表面高表达 CD3+ CD8+ 和 CD3+ CD56+ 双阳性细胞， 与 DCIK 及 CIK 比较差异有统计学意义 （P＜0.05）； CIK、 DCIK、 DCIK-GPC3 培养上清液中 IL-2 生物活性、 浓度以及 IFN-γ 浓度均依次升高， 组间多重比较差异有统计学意义 （P＜0.05）； 在 20∶1 及 50 ∶1 效靶比， CIK、 DCIK、 DCIK- GPC3 对 HepG2 细胞的细胞毒活性依次升高， 组间多重比较差异均有统计学意义 （P＜0.05）。结论 CIK 与 DC- GPC3共培养可获得更强的体外杀伤肝癌细胞活性， 为DCIK-GPC3用于临床免疫治疗提供了理论和实验依据。

Antitumor effects of DC-GPC3 cocultured with CIK on hepatocellular carcinoma in vitro

Abstract: Objective To investigate biological activity and antitumor effects of phosphatidylinositol-3 (GPC3) gene transfected dendritic cells (DC, DC-GPC3) co-cultured with cytokine induced killer cells (CIK) on hepatocellular carcinoma (HCC). Methods The phenotypes of effector cells were analyzed by flow cytometry. Levels of interleukin (IL) - 2 and interferon (IFN)–γ were determined by MTT colorimetry and enzyme-linked immunosorbent assay, respectively. The cytotoxic activity against hepatocarcinoma cells (HepG2) was measured by lactate dehydrogenase release. Results The proportions of CD3+ CD8+ and CD3+ CD56+ double-positive cells were significantly elevated in the DCIK-GPC3 compared with the DCIK and CIK (P＜0.05). Compared with the DCIK and CIK, the DCIK-GPC3 showed significantly higher levels of secreted IL-2 and IFN-γ in the supernatants (P＜0.05). The antitumor effect of DCIK-GPC3 against HepG2 was the highest than that of DCIK and CIK at an effector-target ratio ranging from 20∶1 to 50 ∶1 (P＜0.05). Conclusion DCIK-GPC3 can enhance the cytotoxic activity against hepatocarcinoma cells in vitro. This study provides a theoretical and experimental basis for clinical immunotherapy using DCIK-GPC3.