Tumor cell responses to a novel glutathione S-transferase-activated nitric oxide-releasing prodrug
Tumor cell responses to a novel glutathione S-transferase-activated nitric oxide-releasing prodrug
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DOI：
10.1124/mol.65.5.1070
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发表时间：
2004-05-01
影响因子：
3.6
通讯作者：
Tew, KD
中科院分区：
文献类型：
--
作者：
Findlay, VJ;Townsend, DM;Tew, KD
我们已经使用基于结构的设计技术来引入药物O-2-[2，4-二硝基-5-（N-甲基-N-4-羧基苯基氨基）苯基] 1-N，N-二甲基氨基）二氮烯-1-鎓-1，2-二醇盐（PABA/NO），其通过谷胱甘肽S-转移酶的pi亚型有效代谢为潜在的细胞溶解性一氧化氮，谷胱甘肽S-转移酶是一种在许多肿瘤中高水平表达的酶。我们已经使用了小鼠胚胎成纤维细胞（MEF）空GSTpi（GSTpi（-/-）），以显示GST pi的缺乏导致对PABA/NO的敏感性降低。PABA/NO的细胞毒性也在小鼠皮肤成纤维细胞（NIH 3 T3）细胞系中进行了检查，该细胞系用GSTpi和/或γ-谷氨酰半胱氨酸合成酶和ATP结合盒转运蛋白MRP 1的各种组合稳定转染。MRP 1的过表达赋予最显着程度的阻力，并在体外转运研究证实，一个GSTpi激活的代谢产物PABA/NO被effluxed MRP 1在GSH依赖性的方式。其他研究表明，在缺乏MRP 1的情况下，PABA/NO激活了细胞外调节和应激激活的蛋白激酶ERK，c-Jun NH 2-末端激酶（JNK）和p38。选择性抑制研究表明，JNK和p38的激活是至关重要的PABA/NO的细胞毒性作用。最后，PABA/NO产生的人卵巢癌模型生长在SCID小鼠的抗肿瘤作用。
We have used structure-based design techniques to introduce the drug O-2-[2,4-dinitro-5-(N-methyl-N-4-carboxyphenylamino) phenyl] 1-N,N-dimethylamino) diazen-1-ium-1,2-diolate (PABA/NO), which is efficiently metabolized to potentially cytolytic nitric oxide by the pi isoform of glutathione S-transferase, an enzyme expressed at high levels in many tumors. We have used mouse embryo fibroblasts (MEFs) null for GSTpi (GSTpi(-/-)) to show that the absence of GST pi results in a decreased sensitivity to PABA/NO. Cytotoxicity of PABA/NO was also examined in a mouse skin fibroblast (NIH3T3) cell line that was stably transfected with GSTpi and/or various combinations of gamma-glutamyl cysteine synthetase and the ATP-binding cassette transporter MRP1. Overexpression of MRP1 conferred the most significant degree of resistance, and in vitro transport studies confirmed that a GSTpi-activated metabolite of PABA/NO was effluxed by MRP1 in a GSH-dependent manner. Additional studies showed that in the absence of MRP1, PABA/NO activated the extracellular-regulated and stress-activated protein kinases ERK, c-Jun NH2-terminal kinase (JNK), and p38. Selective inhibition studies showed that the activation of JNK and p38 were critical to the cytotoxic effects of PABA/NO. Finally, PABA/NO produced antitumor effects in a human ovarian cancer model grown in SCID mice.