Depletion of glutathione by buthionine sulfoximine is cytotoxic for human neuroblastoma cell lines via apoptosis
Depletion of glutathione by buthionine sulfoximine is cytotoxic for human neuroblastoma cell lines via apoptosis
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DOI：
10.1006/excr.1998.4303
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发表时间：
1999-01-10
影响因子：
3.7
通讯作者：
Reynolds, CP
中科院分区：
文献类型：
--
作者：
Anderson, CP;Tsai, JM;Reynolds, CP
丁硫基亚砜亚胺（BSO）选择性抑制谷胱甘肽（GSH）的合成，并已被用于敏化肿瘤细胞的烷化剂，但对大多数细胞类型的单一药物的细胞毒性最小。我们使用DIMSCAN（数字图像显微镜细胞毒性试验）测定了BSO对18个（12个MYCN扩增; 6个MYCN非扩增）人神经母细胞瘤细胞系的细胞毒性。D-L（R：S）BSO对大多数神经母细胞瘤细胞系具有高度细胞毒性（>3 log的细胞杀伤），其中17/18个细胞系的IC 90值（范围2.1- >1000 μ M）低于成人人体试验中报道的L（R：S）BSO的等效稳态血浆水平。MYCN基因组扩增的细胞系比MYCN非扩增的细胞系对BSO更敏感（P = 0.04）。D-L（R：S）BSO（500 μ M，72 h）诱导细胞凋亡，如通过DNA梯状、核形态学和使用流式细胞术的DNA片段的TUNEL染色所检测的。最大细胞杀伤发生在48 h内，并且通过添加抗氧化剂（GSH、维生素E和抗坏血酸盐）来拮抗。有趣的是，抗坏血酸具有双峰效应，低剂量逆转和高剂量增强BSO细胞毒性。通过BSO消耗神经母细胞瘤细胞中的GSH导致活性氧（ROS）的形成增加，如通过二氯荧光素二乙酸酯和流式细胞术所测量的。因此，神经母细胞瘤细胞系依赖于GSH作为抗氧化剂来对抗ROS的内源性产生，并且BSO介导的GSH消耗在神经母细胞瘤中可能具有治疗价值，（C）1999 Academic Press。
Buthionine sulfoximine (BSO) selectively inhibits glutathione (GSH) synthesis and has been used to sensitize tumor cells to alkylating agents, but has minimal single-agent cytotoxicity for most cell types. We determined the cytotoxicity of BSO for 18 (12 MYCN amplified; 6 MYCN nonamplified) human neuroblastoma cell lines using DIMSCAN, a digital image microscopy cytotoxicity assay. D-L(R:S) BSO was highly cytotoxic (>3 logs of cell kill) for most neuroblastoma cell lines, with 17/18 cell lines having IC90 values (range 2.1- >1000 mu M) below equivalent steady state plasma levels of L(R:S) BSO reported in adult human trials. Cell lines with genomic amplification of MYCN were more sensitive to BSO than MYCN nonamplified cell lines (P = 0.04). D-L(R:S) BSO (500 mu M for 72 h) induced apoptosis as detected by DNA laddering, nuclear morphology, and TUNEL staining of DNA fragments using flow cytometry, Maximal cell killing occurred within 48 h and was antagonized by the addition of antioxidants (GSH, vitamin E, and ascorbate). Interestingly, ascorbate had a bimodal effect, with lower doses reversing and higher doses enhancing BSO cytotoxicity. Depletion of GSH in neuroblastoma cells by BSO resulted in increased formation of reactive oxygen species (ROS), as measured by dichlorofluorescein diacetate and flow cytometry, Thus, neuroblastoma cell lines rely on GSH as an antioxidant to counter endogenous production of ROS, and BSO-mediated GSH depletion may be of therapeutic value in neuroblastoma, (C) 1999 Academic Press.