GD2特异性单克隆抗体14G2a可诱导IMR-32人神经母细胞瘤细胞凋亡并增强化疗药物的细胞毒性。

The GD2-specific 14G2a monoclonal antibody induces apoptosis and enhances cytotoxicity of chemotherapeutic drugs in IMR-32 human neuroblastoma cells.

作者信息

Kowalczyk Aleksandra, Gil Małgorzata, Horwacik Irena, Odrowaz Zaneta, Kozbor Danuta, Rokita Hanna

机构信息

The Jagiellonian University, Krakow, Poland.

出版信息

Cancer Lett. 2009 Aug 28;281(2):171-82. doi: 10.1016/j.canlet.2009.02.040. Epub 2009 Mar 31.

Abstract

Neuroblastoma (NB) is the most common extracranial solid tumor of childhood. The majority of children suffers from high risk neuroblastoma and has disseminated disease at the time of diagnosis. Despite recent advances in chemotherapy, the prognoses for children with high risk NB remain poor. Therefore, new treatment modalities are urgently needed. GD2 ganglioside is an antigen that is highly expressed on NB cells with only limited distribution on healthy tissues. Consequently, it appears to be an ideal target for both active and passive immunotherapy. The immunological effector mechanisms mediated by anti-GD2 monoclonal antibodies (mAbs) have been already well characterized. However, a growing number of reports suggest that GD2-specific antibodies may exhibit anti-proliferative effects without the immune system involvement. Here, we have shown that anti-GD2 14G2a mAb is capable of decreasing survival of IMR-32 human neuroblastoma cells in a dose-dependent manner. Death induced by this antibody exhibited several characteristics typical for apoptosis such as increased number of Annexin V- and propidium iodide-positive cells, cleavage of caspase 3 and prominent rise in caspase activity. The use of a pan caspase inhibitor Z-VAD-fmk suggested that the killing potential of this mAb is partially caspase-dependent. 14G2a mAb was rapidly endocytosed upon antigen binding. Employment of chloroquine, an inhibitor of lysosomal degradation, did not rescue IMR-32 cells from antibody-induced cell death suggesting lack of ceramide involvement in the observed effect. Most importantly, our studies showed that at particular drug concentrations 14G2a mAb exerts a synergistic effect with doxorubicin and topotecan, as well as an additive effect with carboplatin in killing IMR-32 cells in vitro. Our results provide guidance regarding how to best combine GD2-specific 14G2a antibody with existing cancer therapeutic agents to improve available treatment modalities for neuroblastoma.

摘要

神经母细胞瘤(NB)是儿童最常见的颅外实体瘤。大多数儿童患有高危神经母细胞瘤,在诊断时已出现播散性疾病。尽管化疗最近取得了进展,但高危NB患儿的预后仍然很差。因此,迫切需要新的治疗方法。GD2神经节苷脂是一种在NB细胞上高度表达而在健康组织中分布有限的抗原。因此,它似乎是主动和被动免疫治疗的理想靶点。由抗GD2单克隆抗体(mAb)介导的免疫效应机制已经得到了很好的表征。然而,越来越多的报告表明,GD2特异性抗体可能在没有免疫系统参与的情况下表现出抗增殖作用。在这里,我们已经表明抗GD2 14G2a单克隆抗体能够以剂量依赖的方式降低IMR-32人神经母细胞瘤细胞的存活率。这种抗体诱导的死亡表现出凋亡的几个典型特征,如膜联蛋白V和碘化丙啶阳性细胞数量增加、半胱天冬酶3的切割和半胱天冬酶活性的显著升高。使用泛半胱天冬酶抑制剂Z-VAD-fmk表明,这种单克隆抗体的杀伤潜力部分依赖于半胱天冬酶。14G2a单克隆抗体在抗原结合后迅速被内吞。使用溶酶体降解抑制剂氯喹并不能使IMR-32细胞从抗体诱导的细胞死亡中获救,这表明观察到的效应中缺乏神经酰胺的参与。最重要的是,我们的研究表明,在特定的药物浓度下,14G2a单克隆抗体与阿霉素和拓扑替康具有协同作用,与卡铂在体外杀死IMR-32细胞时具有相加作用。我们的结果为如何最好地将GD2特异性14G2a抗体与现有的癌症治疗药物联合使用以改善神经母细胞瘤的现有治疗方法提供了指导。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索