Synergistic cytotoxicity in solid tumor cell lines between N-(4-hydroxyphenyl)retinamide and modulators of ceramide metabolism
Synergistic cytotoxicity in solid tumor cell lines between N-(4-hydroxyphenyl)retinamide and modulators of ceramide metabolism
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DOI：
10.1093/jnci/92.23.1897
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发表时间：
2000-12-06
影响因子：
10.3
通讯作者：
Reynolds, CP
中科院分区：
文献类型：
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作者：
Maurer, BJ;Melton, L;Reynolds, CP
背景资料：我们以前报道过，N-（4-羟基苯基）维A酰胺（4-HPR，芬维A胺）治疗引起神经母细胞瘤细胞系中神经酰胺水平的大幅增加，并通过p53（也称为TP 53）非依赖性和半胱天冬酶非依赖性途径诱导细胞凋亡和坏死。我们的目标是确定几种抑制神经酰胺代谢酶的分子-L-苏型-二氢鞘氨醇（safingol）、d，l-苏型-1-苯基-2-十六烷酰氨基-3-吗啉代-1-丙醇（PPMP）和他莫昔芬是否增强4-HPR介导的细胞毒性和/或影响神经酰胺水平。方法：采用放射性标记和薄层色谱法测定细胞脂质。使用数字成像显微镜，使用基于荧光的细胞活力测定法，在培养的癌细胞系中测量细胞毒性和细胞毒性协同作用（表示为组合指数，其中组合指数1表示拮抗作用）。统计检验是双侧的。结果如下：4-HPR通过de nova合成增加神经酰胺水平，将Safingol（1-4 μ M）掺入神经酰胺的立体化学变体中，并与3：1摩尔比的4-HPR（3-12 μ M）协同作用，在神经母细胞瘤中产生相对于单独的4-HPR 100倍至10000倍（2至4 log）的细胞毒性增加（组合指数
Background: We previously reported that N-(4-hydroxyphenyl)retinamide (4-HPR, fenretinide) treatment caused large increases of ceramide levels in neuroblastoma cell lines and induced cell death by a combination of apoptosis and necrosis through p53 (also known as TP53)-independent and caspase-independent pathways. Our goal was to determine if several molecules that inhibit enzymes involved in ceramide metabolism-L-threo-dihydrosphingosine (safingol), d,l-threo-1-phenyl-2-hexadecanoylamino-3-morpholino-1-propanol (PPMP), and tamoxifen-enhanced 4-HPR-mediated cytotoxicity and/or affected ceramide levels. Methods: Cellular lipids were quantified by radiolabeling and thin-layer chromatography. Cytotoxicity and cytotoxic synergy (expressed as combination index, where combination index 1 indicates antagonism) were measured in cultured cancer cell lines with the use of a fluorescence-based assay of cen viability employing digital imaging microscopy, Statistical tests were two-sided. Results: 4-HPR increased ceramide levels by de nova synthesis, Safingol (1-4 muM) was incorporated into a stereochemical variant of ceramide and synergized with a 3:1 molar ratio of 4-HPR (3-12 muM), to produce a 100-fold to 10 000-fold (2 to 4 logs) increase in cytotoxicity relative to 4-HPR alone in neuroblastoma (combination index