Synergistic Effects of Oncolytic Reovirus and Cisplatin Chemotherapy in Murine Malignant Melanoma
Synergistic Effects of Oncolytic Reovirus and Cisplatin Chemotherapy in Murine Malignant Melanoma
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DOI：
10.1158/1078-0432.ccr-09-0796
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发表时间：
2009-10-01
影响因子：
11.5
通讯作者：
Morgan, Richard
中科院分区：
文献类型：
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作者：
Pandha, Hardev S.;Heinemann, Lucy;Morgan, Richard
目的：在人、小鼠黑色素瘤细胞株和小鼠黑色素瘤模型中检测呼肠孤病毒联合顺铂化疗方案，并探讨其协同抗肿瘤作用的可能机制。实验设计：采用3-(4，5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium法和空斑试验评价呼肠孤病毒+/-化疗对体外细胞毒作用和病毒复制的影响。用组合指数分析评价药物间的相互作用。采用Annexin V/Propidium iodide荧光激活的细胞分选方法评价细胞死亡模式；采用Agilent微阵列系统完成单一处理和联合处理的基因表达谱分析。结果：活体呼肠孤病毒与多种化疗药物在B16-F10小鼠黑色素瘤细胞中有不同程度的协同细胞毒作用，其中以顺铂的协同作用最为明显(ED(50)时联合指数为0.42+/-0.03)。顺铂和呼肠孤病毒的联合暴露导致晚期凋亡/坏死细胞数量增加。顺铂几乎完全阻断呼肠孤病毒诱导的炎性细胞因子基因上调。联合治疗显著延缓了肿瘤生长，提高了体内存活率(P<；0.0001和P=0.0003)。顺铂对呼肠孤病毒引起的小鼠体液反应无影响。结论：呼肠孤病毒与多种化疗药物联合应用，可协同增强人黑色素瘤细胞株、小鼠黑色素瘤细胞株和小鼠黑色素瘤细胞的细胞毒作用。这些数据支持目前正在临床上进行的呼肠孤病毒/化疗组合I期临床研究。(临床癌症研究2009；15(19)：6158-66)
Purpose: To test combination treatment schedules of reovirus and cisplatin chemotherapy in human and murine melanoma cell lines and murine models of melanoma and to investigate the possible mechanisms of synergistic antitumor effects.Experimental Design: The effects of reovirus +/- chemotherapy on in vitro cytotoxicity and viral replication were assessed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay and plaque assay. Interactions between agents were assessed by combination index analysis. Mode of cell death was assessed by Annexin V/propidium iodide fluorescence-activated cell sorting-based assays; gene expression profiling of single versus combination treatments was completed using the Agilent microarray system. Single agent and combination therapy effects were tested in vivo in two immunocompetent models of murine melanoma.Results: Variable degrees of synergistic cytotoxicity between live reovirus and several chemotherapy agents were observed in B16.F10 mouse melanoma cells, most significantly with cisplatin (combination index of 0.42 +/- 0.03 at ED(50)). Combination of cisplatin and reovirus exposure led to increased late apoptotic/necrotic cell populations. Cisplatin almost completely abrogated the inflammatory cytokine gene up-regulation induced by reovirus. Combination therapy led to significantly delayed tumor growth and improved survival in vivo (P < 0.0001 and P = 0.0003, respectively). Cisplatin had no effect on the humoral response to reovirus in mice. However, cisplatin treatment suppressed the cytokine and chemokine response to reovirus in vitro and in vivo.Conclusion: The combination of reovirus and several chemotherapeutic agents synergistically enhanced cytotoxicity in human and murine melanoma cell lines in vitro and murine tumors in vivo. The data support the current reovirus/chemotherapy combination phase I clinical studies currently ongoing in the clinic. (Clin Cancer Res 2009; 15(19):6158-66)