Sindbis viral structural protein cytotoxicity on human neuroblastoma cells.
Sindbis viral structural protein cytotoxicity on human neuroblastoma cells.
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辛德比斯病毒结构蛋白对人神经母细胞瘤细胞的细胞毒性。
DOI：
10.1007/s00383-020-04719-8
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Yoshida H.
中科院分区：
文献类型：
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作者：
Saito EY;Saito K;Hishiki T;Takenouchi A;Saito T;Sato Y;Terui K;Matsunaga T;Shirasawa H;Yoshida H.
目的辛德毕斯病毒 (SINV) 对神经母细胞瘤 (NB) 细胞进行溶瘤病毒治疗是治疗高危 NB 的一种有前途的策略。在这里，我们评估了使用 SINV 结构蛋白作为 NB 治疗剂的可能性，因为紫外线灭活的 SINV 可能诱导细胞病变效应。方法紫外线灭活的 SINV 对人 NB 细胞系 NB69、NGP、GOTO、NLF、SK-N-SH、SH-SY5Y、CHP134、NB-1、IMR32 和 分析了RT-BM-1。通过TUNEL 测定证实细胞凋亡。为了确定负责 UV 灭活 SINV 细胞毒性的 SINV 成分，将编码结构蛋白（即衣壳、E2 和 E1）的表达载体转染到 NB 细胞中。通过MTT测定评估细胞毒性。结果UV灭活的SINV在NB69、NGP和RT-BM-1中比在正常人成纤维细胞中引发更显着的细胞毒性。转染实验结果表明，所有对紫外线灭活的SINV敏感的NB细胞系都对E1蛋白高度敏感，而转染带有衣壳、E1或E2的载体的成纤维细胞则不然。 可选择性地用作NB的治疗剂。
PurposeOncolytic viral therapy for neuroblastoma (NB) cells with Sindbis virus (SINV) is a promising strategy for treating high-risk NB. Here, we evaluated the possibility of using SINV structural proteins as therapeutic agents for NB since UV-inactivated SINV could induce cytopathogenic effects.MethodsThe cytotoxicity of UV-inactivated SINV toward human NB cell lines NB69, NGP, GOTO, NLF, SK-N-SH, SH-SY5Y, CHP134, NB-1, IMR32, and RT-BM-1 were analyzed. Apoptosis was confirmed by TUNEL assays. To determine the components of SINV responsible for the cytotoxicity of UV-inactivated SINV, expression vectors encoding the structural proteins, namely capsid, E2, and E1, were transfected in NB cells. Cytotoxicity was evaluated by MTT assays.ResultsUV-inactivated SINV elicited more significant cytotoxicity in NB69, NGP, and RT-BM-1 than in normal human fibroblasts. Results of the transfection experiments showed that all NB cell lines susceptible to UV-inactivated SINV were highly susceptible to the E1 protein, whereas fibroblasts transfected with vectors harboring capsid, E1, or E2 were not.ConclusionsWe demonstrated that the cytotoxicity of the UV-inactivated SINV is due to apoptosis induced by the E1 structural protein of SINV, which can be used selectively as a therapeutic agent for NB.