Allelic variants of the human glutathione S-transferase P1 gene confer differential cytoprotection against anticancer agents in Escherichia coli
Allelic variants of the human glutathione S-transferase P1 gene confer differential cytoprotection against anticancer agents in Escherichia coli
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DOI：
10.1097/00008571-200210000-00006
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发表时间：
2002-10-01
期刊：
影响因子：
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通讯作者：
Ali-Osman, F
中科院分区：
文献类型：
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作者：
Ishimoto, TM;Ali-Osman, F
多态的人类GSTP1基因位点编码差异代谢亲电底物的蛋白质，包括临床癌症治疗中使用的许多化疗药物。在本研究中，我们利用大肠杆菌XL1-Blue MRF’菌株，用携带三个GSTP1等位基因cdna的噬菌体表达载体转化，研究了不同GSTP1等位基因对卡铂、顺铂、硫替帕和4-氢过氧异环磷酰胺四种临床活性抗癌药物的细胞保护能力。异丙基p- d -硫代半乳糖苷诱导蛋白表达后，分别用每种药物处理细胞3小时（4-氢过氧异环磷酰胺处理11小时）。幸存的部分被确定并用于计算每个等位基因对每种药物的细胞保护因子。结果表明，所有GSTP1等位基因都具有细胞保护作用，尽管程度不同。对于顺铂和卡铂，GSTP1* C等位基因的保护作用最强，Cps分别为5.58和3.76，而GSTP1* A和GSTP1*B的Cps分别为1.21和1.61和2.50和2.79。相比之下，GSTP1*A对硫替帕的保护作用最高，其细胞保护因子为1.56，而GSTP1*B和GSTP1*C的细胞保护因子分别为1.32和1.1。对于4-氢过氧异环磷酰胺，GSTP1*B和GSTP1*C的细胞保护因子相同，为1.45，而GSTP1*A的细胞保护因子为1.18。这些数据表明，不同GSTP1等位基因对亲电抗癌药物细胞毒性的保护能力存在显著差异。不同的GSTP1等位基因和不同的抗癌药物之间的保护水平存在显著差异。所描述的优化的原核系统为基因对药物敏感性的影响的药物遗传学分析提供了一个有用和快速的工具。
The polymorphic human GSTP1 gene locus encodes proteins that differentially metabolize electrophilic substrates, including, many chemotherapeutic agents used in clinical cancer therapy. In this study, we used Escherichia coli XL1-Blue MRF' strain, transformed with phagemid expression vectors carrying cDNAs of three GSTP1 alleles, to investigate the cytoprotective abilities of the different GSTP1 alleles against four clinically active anticancer agents, namely, carboplatin, cisplatin, thiotepa, and 4-hydroperoxyifosfamide. Following induction of protein expression with isopropyl-p-D-thiogalactoside, the cells were treated with each drug for 3 h (11 h for 4-hydroperoxyifosfamide). Surviving fractions were determined and used to compute a cytoprotective factor for each allele against each drug. The results showed all the GSTP1 alleles to be cytoprotective, albeit to different degrees. For cisplatin and carboplatin, the GSTP1* C allele was most protective, with Cps of 5.58 and 3.76, respectively, compared with 1.21 and 1.61 for GSTP1 *A and 2.50 and 2.79 for GSTP1*B. In contrast protection against thiotepa was highest for the GSTP1*A allele, with a cytoprotective factor of 1.56, compared to 1.32 for GSTP1*B and 1.1 for GSTP1* C. For 4-hydroperoxyifosfamide, the CP for GSTP1*B and GSTP1*C was the same, 1.45, compared with 1.18 for GSTP1 *A. These data demonstrate significant differences in the ability of the different GSTP1 alleles to protect against the cytotoxicity of electrophilic anticancer agents. The level of protection differs significantly between different GSTP1 alleles, and between different anticancer agents. The optimized prokaryotic system described provides a useful and rapid tool for pharmacogenetic analysis of the effects of genes on drug sensitivity.