DNA-protein crosslink and DNA strand break formation in HL-60 cells treated with trans,trans-muconaldehyde, hydroquinone and their mixtures.
DNA-protein crosslink and DNA strand break formation in HL-60 cells treated with trans,trans-muconaldehyde, hydroquinone and their mixtures.
复制标题
用反式、反式粘康醛、氢醌及其混合物处理的 HL-60 细胞中 DNA-蛋白质交联和 DNA 链断裂形成。
DOI：
10.1080/10915810151115173
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发表时间：
2001
影响因子：
2.2
通讯作者：
Witz,G
中科院分区：
文献类型：
--
作者：
Amin,RP;Witz,G
苯是一种人类白血病原，也是一种普遍存在的环境污染物，其毒性部分是由包括对苯二酚(HQ)在内的环羟化代谢产物和包括反式、反式邻苯二甲醛(MUC)在内的开环代谢产物及其相互作用所介导的。DNAPC和DNAASB是与致癌化合物毒性机制(S)相关的毒性损伤。在本研究中，我们检验了假设，即MUC和HQ的个体效应和交互作用参与了DNAPC和DNASB的形成。我们将以前关于MUC诱导HL-60细胞DNAPC的研究扩展到HQ以及MUC和HQ的混合物，并测定了DNASB的水平，包括3‘OH DNASB。K+/十二烷基硫酸钠沉淀法显示，HL-60细胞经25~100μM-HQ处理4小时后，DNA-PC水平较对照组增加1.3~2.8倍。在25和100μM时，MUC诱导DNAPC的效率分别是HQ的1.8倍和4.9倍。用25或50μ、M、MUC和HQ的等摩尔混合物处理，相对于如果作用只是相加的情况下预期的DNAPC水平，导致更高的DNAPC形成。TUNEL法检测显示，5~25μMUC作用1h后，3‘OHDNASB水平呈浓度依赖性增加，而HQ处理则无明显影响。与单独使用MUC相比，与25和50μM MUC/HQ混合物共同处理导致TUNEL标记显着减少。流式细胞仪检测显示，经1-50μM MUC或HQ处理的HL-60细胞的DNASB呈浓度和时间依赖性增加，该实验测量了各种单链和双链断裂以及碱不稳定位点。暴露于10μM MUC的1小时和2小时时间点的Qdnasb值(1Qdnasb≍100DNASB/cell)分别为7.5±1.2和15.4±1.4，而相应时间对照组分别为0.1±3.8和0.0±1.5。10μMHQ治疗后1小时和2小时QdNasb值分别为4.4±0.7和17.7±2.1，而相应时间对照组分别为0.0±0.5和0.0±1.3。5~50μM等摩尔的MUC/HQ混合液作用1小时后，DNASB的诱导是相加的。这些体外结果对于MUC和HQ诱导的DNAPC和DNASB的损伤以及它们之间的相互作用可能有助于苯诱导的血液毒性和白血病的发生具有重要意义。
The toxicity of benzene, a human leukemogen and ubiquitous environmental pollutant, is mediated in part by ring-hydroxylated metabolites including hydroquinone (HQ) and ring-opened metabolites includingtrans,trans-muconaldehyde (muconaldehyde, MUC), and their interactions. DNA-protein crosslinks (DNAPC) and DNA strand breaks (DNASB) are toxic lesions associated with the mechanism(s) of toxicity of carcinogenic compounds. In the present studies, we examined the hypothesis that individual and interactive effects of MUC and HQ are involved in the formation of DNAPC and DNASB. We extended our previous studies on DNAPC induction by MUC in HL-60 cells to HQ and mixtures of MUC and HQ, and determined DNASB levels, including 3'OH DNASB. Treatment of HL-60 cells with 25 to 100 μM HQ followed by incubation for 4 hours resulted in 1.3- to 2.8-fold increases in DNAPC levels compared with control, as determined by a K+/sodium dodecyl sulfate (SDS) precipitation assay. At 25 and 100 μM, MUC was 1.8 and 4.9 fold more effective at inducing DNAPC than HQ. Treatment with equimolar mixtures of 25 or 50 μM MUC and HQ resulted in higher DNAPC formation relative to the DNAPC levels expected if the effects were only additive. 3'OH DNASB levels as determined by the TUNEL assay showed a significant concentration-dependent increase 1 hour after treatment with 5 to 25 μM MUC, whereas HQ treatment had no effect. Cotreatment with 25 and 50 μM MUC/HQ mixtures resulted in significant decreases in TUNEL labeling relative to treatment with MUC alone. HL-60 cells treated with 1 to 50 μM MUC or HQ exhibited concentration- and time-dependent increases in DNASB as determined by the FADU assay, which measures a variety of single- and double-strand breaks and alkali labile sites. Exposure to 10 μM MUC gaveQdnasbvalues (1Qdnasb≍100 DNASB/cell) of 7.5 ± 1.2 and 15.4 ± 1.4 at the 1- and 2-hour time points respectively, compared with 0.1 ± 3.8 and 0.0 ± 1.5 for the corresponding time controls. TheQdnasbvalues after treatment with 10 μM HQ were 4.4 ± 0.7 and 17.7±2.1 at the 1- and 2-hour time points, respectively, compared with 0.0 ± 0.5 and 0.0 ± 1.3 for the corresponding time controls. Induction of DNASB was additive 1 hour after treatment with equimolar MUC/HQ mixtures of 5 to 50 μM. These in vitro findings are significant in that DNAPC and DNASB lesions induced by MUC and HQ as well as their interactions could contribute to benzene-induced hematotoxicity and leukemogenesis.