DC-GPC3联合CIK细胞的体外抗肝癌作用

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

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。

Objective To investigate biological activity and antitumor effects of phosphatidylinositol-3(GPC3) genetransfected 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 andinterferon(IFN) – γ were determined by MTT colorimetry and enzyme-linked immunosorbent assay, respectively. Thecytotoxic activity against hepatocarcinoma cells(HepG2) was measured by lactate dehydrogenase release. Results Theproportions of CD3^+CD8^+and CD3^+CD56^+double-positive cells were significantly elevated in the DCIK-GPC3 comparedwith the DCIK and CIK(P<0.05). Compared with the DCIK and CIK, the DCIK-GPC3 showed significantly higher levels ofsecreted IL-2 and IFN-γ in the supernatants(P<0.05). The antitumor effect of DCIK-GPC3 against HepG2 was the highestthan that of DCIK and CIK at an effector-target ratio ranging from 20∶1 to 50∶1(P<0.05). Conclusion DCIK-GPC3 canenhance the cytotoxic activity against hepatocarcinoma cells in vitro. This study provides a theoretical and experimental basisfor clinical immunotherapy using DCIK-GPC3.