Persistent inhibition of FLIPL expression by lentiviral small hairpin RNA delivery restores death-receptor-induced apoptosis in neuroblastoma cells
Persistent inhibition of FLIPL expression by lentiviral small hairpin RNA delivery restores death-receptor-induced apoptosis in neuroblastoma cells
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DOI：
10.1007/s10495-006-3435-9
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发表时间：
2006-02-01
期刊：
影响因子：
7.2
通讯作者：
Gross, N
中科院分区：
文献类型：
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作者：
Flahaut, M;Mühlethaler-Mottet, A;Gross, N
神经母细胞瘤是儿童期最常见和最致命的实体瘤，其不同的生物学和临床行为可以通过细胞凋亡的差异调节来解释。为了了解神经母细胞瘤细胞中死亡抵抗的潜在机制，我们开发了由慢病毒载体产生的RNA小发夹作为选择性干扰FLIPL的工具，FLIPL是死亡受体诱导的细胞凋亡的主要负调节因子。这些工具在干扰FLIPL表达和功能方面表现出高度有效性，因为它们几乎完全抑制内源性和/或外源性过表达的FLIPL蛋白，并完全逆转FLIPL介导的TRAIL抗性。此外，干扰内源性FLIPL和FLIPS显着恢复FasL的敏感性在SH-EP神经母细胞瘤细胞系。这些结果揭示了慢病毒介导的shRNA特异性和持续干扰FLIP表达的能力，并支持FLIP参与神经母细胞瘤细胞中死亡受体介导的凋亡的调节。将这些工具与其他治疗方式相结合可能会改善对神经母细胞瘤等耐药肿瘤的治疗。
Neuroblastoma represents the most common and deadly solid tumour of childhood, which disparate biological and clinical behaviour can be explained by differential regulation of apoptosis. To understand mechanisms underlying death resistance in neuroblastoma cells, we developed small hairpin of RNA produced by lentiviral vectors as tools to selectively interfere with FLIPL, a major negative regulator of death receptor-induced apoptosis. Such tools revealed highly efficient in interfering with FLIPL expression and function as they almost completely repressed endogenous and/or exogenously overexpressed FLIPL protein and fully reversed FLIPL-mediated TRAIL resistance. Moreover, interference with endogenous FLIPL and FLIPS significantly restored FasL sensitivity in SH-EP neuroblastoma cell line. These results reveal the ability of lentivirus-mediated shRNAs to specifically and persistently interfere with FLIP expression and support involvement of FLIP in the regulation of death receptor-mediated apoptosis in neuroblastoma cells. Combining such tools with other therapeutic modalities may improve treatment of resistant tumours such as neuroblastoma.