Enhanced glutathione depletion, protein adduct formation, and cytotoxicity following exposure to 4-hydroxy-2-nonenal (HNE) in cells expressing human multidrug resistance protein-1 (MRP1) together with human glutathione S-transferase-M1 (GSTM1).
Enhanced glutathione depletion, protein adduct formation, and cytotoxicity following exposure to 4-hydroxy-2-nonenal (HNE) in cells expressing human multidrug resistance protein-1 (MRP1) together with human glutathione S-transferase-M1 (GSTM1).
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DOI：
10.1016/j.cbi.2011.08.012
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发表时间：
2011-11-15
影响因子：
5.1
通讯作者：
Townsend, Alan J.
中科院分区：
文献类型：
--
作者：
Rudd, Lisa P.;Kabler, Sandra L.;Morrow, Charles S.;Townsend, Alan J.
关键词：
4-羟基-2-壬烯醛（HNE）是脂质过氧化反应最活跃的产物之一，在细胞中具有细胞毒性和遗传毒性作用。已经报道了几种酶促途径使HNE解毒，包括通过谷胱甘肽-S-转移酶（GST）的缀合。完全解毒可能需要通过外排转运蛋白去除产生的HNE-谷胱甘肽结合物（HNE-SG）。我们研究了GSTM 1和/或ABC外排转运蛋白，多药耐药蛋白-1（MRP 1）的表达对HNE诱导的细胞毒性的影响。稳定转染的MCF 7细胞系用于检查GSTM 1和/或MRP 1表达对HNE诱导的细胞毒性、GSH消耗和HNE-蛋白加合物形成的影响。在MCF 7细胞系中GSTM 1与MRP 1的共表达导致对HNE细胞毒性的2.3倍致敏作用（相对于对照的0.44倍IC 50值），而不是预期的保护作用。单独表达GSTM 1或MRP 1也导致对HNE细胞毒性的轻微致敏（IC 50值分别降低0.79倍和0.71倍）。相对于不表达对照细胞系，GSTM 1和MRP 1的共表达强烈增强了HNE-蛋白加合物的形成，而单独MRP 1或单独GSTM 1的表达产生与对照细胞系类似的低水平的HNE-蛋白加合物。在对照细胞系或MCF 7/GSTM 1细胞系中，用相同的HNE暴露60分钟，谷胱甘肽（GSH）水平降低10-20%。然而，HNE在单独表达MRP 1的细胞中诱导> 80%的GSH消耗。MRP 1和GSTM 1的共表达引起稍微更大的GSH消耗，与该细胞系中更大的蛋白加合物形成和细胞毒性一致。由于单独表达GSTM 1或MRP 1不会使细胞对HNE强烈敏感，或导致比对照细胞系中更多的HNE-蛋白加合物，因此这些结果表明MRP 1和GSTM 1协同增强HNE-蛋白加合物形成和HNE细胞毒性，这是由MRP 1和GSTM 1两者介导的GSH耗竭促进的。
4-hydroxy-2-nonenal (HNE) is one of the most reactive products of lipid peroxidation and has both cytotoxic and genotoxic effects in cells. Several enzymatic pathways have been reported to detoxify HNE, including conjugation by glutathione-S-transferases (GSTs). Removal of the resulting HNE-glutathione conjugate (HNE-SG) by an efflux transporter may required for complete detoxification. We investigated the effect of expression of GSTM1 and/or the ABC efflux transporter protein, multidrug-resistance protein-1 (MRP1), on HNE-induced cellular toxicity. Stably transfected MCF7 cell lines were used to examine the effect of GSTM1 and/or MRP1 expression on HNE-induced cytotoxicity, GSH depletion, and HNE-protein adduct formation. Co-expression in the MCF7 cell line of GSTM1 with MRP1 resulted in a 2.3-fold sensitization to HNE cytotoxicity (0.44-fold IC50 value relative to control) rather than the expected protection. Expression of either GSTM1 or MRP1 alone also resulted in slight sensitization to HNE cytotoxicity (0.79-fold and 0.71-fold decreases in IC50 values, respectively). Co-expression of GSTM1 and MRP1 strongly enhanced the formation of HNE-protein adducts relative to the non-expressing control cell line, whereas expression of either MRP1 alone or GSTM1 alone yielded similarly low levels of HNE-protein adducts to that of the control cell line. Glutathione (GSH) levels were reduced by 10–20% in either the control cell line or the MCF7/GSTM1 cell line with the same HNE exposure for 60 minutes. However, HNE induced > 80% depletion of GSH in cells expressing MRP1 alone. Co-expression of both MRP1 and GSTM1 caused slightly greater GSH depletion, consistent with the greater protein adduct formation and cytotoxicity in this cell line. Since expression of GSTM1 or MRP1 alone did not strongly sensitize cells to HNE, or result in greater HNE-protein adducts than in the control cell line, these results indicate that MRP1 and GSTM1 collaborate to enhance HNE-protein adduct formation and HNE cytotoxicity, facilitated by GSH depletion mediated by both MRP1 and GSTM1.
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影响因子：
4.1
作者：
Hubatsch, I;Ridderström, M;Mannervik, B
通讯作者：
Mannervik, B
影响因子：
3.8
作者：
Knight, Tamara Raphael;Choudhuri, Supratim;Klaassen, Curtis D.
通讯作者：
Klaassen, Curtis D.
影响因子：
3.8
作者：
Diah, SK;Smitherman, PK;Morrow, CS
通讯作者：
Morrow, CS
DOI：
10.1093/jnci/85.14.1159
发表时间：
1993-07-21
期刊：
JOURNAL OF THE NATIONAL CANCER INSTITUTE
影响因子：
--
作者：
BELL, DA;TAYLOR, JA;LUCIER, GW
通讯作者：
LUCIER, GW
影响因子：
7.4
作者：
Ji, B;Ito, K;Horie, T
通讯作者：
Horie, T