热休克蛋白70联合白介素--2对小鼠肝癌细胞株移植瘤的影响

Collaborative effect of tumor-deri ved HSP70 and IL-2 on mouse hepatocellular carcinoma cell line implanted tumor

目的:研究肿瘤热休克蛋白70联合应用IL-2对小鼠肝癌的治疗作用,为两种免疫制剂联合应用治疗人类恶性肿瘤提供参考依据。方法:应用细胞培养、蛋白提纯、电泳、Western-blot、毛细管电泳、动物实验等方法。小鼠分别用HSP70 10μg、5μg和IL-2 50kU、100kU、2kU(维持量)按设计的方案进行治疗试验。结果:单纯应用或联合应用HSP70和IL-2均能明显地延缓小鼠移植肝癌的生长并能不同程度地延长生存期,与对照组相比(除IL-250kU组外)差异显著(P<0.025-0.05)。但获长期存活(>90d)者仅见于接受HSP70 10μg治疗组,40％(2/5)的HSP70 10μg组及HSP70 10μg联合应用IL-2 50kU组小鼠,和60％(3/5)的HSP70 10μg联合应用IL-2 100kU组小鼠,获长期生存的小鼠其肿瘤最终全部消退。与无长期生存小鼠的其他组相比差异十分显著(P<0.025-0.05)。IL-2 100kU与HSP70 5μg的作用相当。结论:肿瘤来源的HSP70较IL-2具有更强的治疗作用,在联合应用实验的治疗组中HSP70对延缓和消除肿瘤起主要作用。更重要的是,合适剂量的HSP70与IL-2联合应用(本研究中HSP70 10μg与IL-2 100kU)能明显地提高对荷瘤小鼠的治疗作用。

AIM: To investigate the therapeutic efficacy of compound immunotherapy of tumor-derived HSP70 and IL-2 on mouse hepatocellular carcinoma model. METHODS: Cell-culture, techniques for protein extraction and purification, SDS-PAGE, Western-blot, and animal experiment were applied: in this study. Mice were treated with HSP70 10μg, 5μg and IL-2 50kU, 100kU and 2kU (maintaining dosage) at previously designated interval. RESULTS: HSP70 or IL-2, and the compound imrnunotherapy of HSP70 and IL-2 have obviously delayed the growth of the implanted tumor and prolonged life span of the mice in different extent, and significant differences were observed (except IL-2 50kU group), compared to control group (P<0.025-0.05). Long period tumor-free survival (over 90 days) rate was found merely in the groups received HSP70 10μg, 40％ (2/5) in HSP70 10μg group and HSP70 10μg-IL-2 50kU group, and 60％ (3/5) in HSP70 10μg-IL-2 100 kU group, whereas none of the mice in the rest groups obtained a long period survival. Statistics significance was apparent compared with the groups without long period survival mice (P<0.025-0.05). IL-2 100kU administration was as effective as HSP70 5μJg. CONCLUSIONS: Tumor-derived HSP70 immunotherapy demonstrates much more therapeutic efficacy than IL-2, and compound immunotherapy HSP70 exerts main activity in delaying or eradicating the tumor. Importantly, the combination of proper dosage of HSP70 and IL-2 (HSP70 10μg and IL-2 100kU) has obviously improved the immunotherapy efficacy, which may be a promising strategy to treat human cancer.