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暴露于致癌多环芳烃的大鼠肝上皮“干细胞样”细胞中DNA加合物的形成及细胞凋亡的诱导

DNA adducts formation and induction of apoptosis in rat liver epithelial 'stem-like' cells exposed to carcinogenic polycyclic aromatic hydrocarbons.

作者信息

Topinka Jan, Marvanová Sona, Vondrácek Jan, Sevastyanova Oksana, Nováková Zuzana, Krcmár Pavel, Pencíková Katerina, Machala Miroslav

机构信息

Laboratory of Genetic Ecotoxicology, Institute of Experimental Medicine, AS CR, 142 20 Prague, Czech Republic.

出版信息

Mutat Res. 2008 Feb 1;638(1-2):122-32. doi: 10.1016/j.mrfmmm.2007.09.004. Epub 2007 Sep 16.

Abstract

The bipotent liver progenitor cells, so called oval cells, may participate at the early stages of hepatocarcinogenesis induced by chemical carcinogens. Unlike in mature parenchymal cells, little is known about formation of DNA adducts and other genotoxic events in oval cells. In the present study, we employed spontaneously immortalized rat liver WB-F344 cell line, which is an established in vitro model of oval cells, in order to study genotoxic effects of selected carcinogenic polycyclic aromatic hydrocarbons (PAHs). With exception of dibenzo[a,l]pyrene, and partly also benzo[g]chrysene and benz[a]anthracene, all other PAHs under the study induced high levels of CYP1A1 and CYP1B1 mRNA. In contrast, we observed distinct genotoxic and cytotoxic potencies of PAHs. Dibenzo[a,l]pyrene, and to a lesser extent also benzo[a]pyrene, benzo[g]chrysene and dibenzo[a,e]pyrene, formed high levels of DNA adducts. This was accompanied with accumulation of Ser-15 phosphorylated form of p53 protein and induction of apoptosis. Contrary to that, benz[a]anthracene, chrysene, benzo[b]fluoranthene and dibenzo[a,h]anthracene induced only low amounts of DNA adducts formation and minimal apoptosis, without exerting significant effects on p53 phosphorylation. Finally, we studied effects of 2,4,3',5'-tetramethoxystilbene and fluoranthene, inhibitors of CYP1B1 activity, which plays a central role in metabolic activation of dibenzo[a,l]pyrene. In a dose-dependent manner, both compounds inhibited apoptosis induced by dibenzo[a,l]pyrene, suggesting that it interferes with the metabolic activation of the latter one. The present data show that in model cell line sharing phenotypic properties with oval cells, PAHs can be efficiently metabolized to form ultimate genotoxic metabolites. Liver progenitor cells could be thus susceptible to this type of genotoxic insult, which makes WB-F344 cell line a useful tool for studies of genotoxic effects of organic contaminants in liver cells. Our results also suggest that, unlike in mature hepatocytes, CYP1B1 might be a primary enzyme responsible for formation of DNA adducts in liver progenitor cells.

摘要

双能肝祖细胞,即所谓的卵圆细胞,可能参与化学致癌物诱导的肝癌发生早期阶段。与成熟实质细胞不同,关于卵圆细胞中DNA加合物的形成及其他基因毒性事件知之甚少。在本研究中,我们采用自发永生化大鼠肝WB - F344细胞系,这是一种已建立的卵圆细胞体外模型,以研究选定的致癌多环芳烃(PAHs)的基因毒性作用。除二苯并[a,l]芘外,部分苯并[g] Chrysene和苯并[a]蒽也除外,研究中的所有其他PAHs均诱导高水平的CYP1A1和CYP1B1 mRNA。相比之下,我们观察到PAHs具有明显不同的基因毒性和细胞毒性。二苯并[a,l]芘,以及程度较轻的苯并[a]芘、苯并[g] Chrysene和二苯并[a,e]芘,形成高水平的DNA加合物。这伴随着p53蛋白Ser - 15磷酸化形式的积累和细胞凋亡的诱导。与此相反,苯并[a]蒽、 Chrysene、苯并[b]荧蒽和二苯并[a,h]蒽仅诱导少量的DNA加合物形成和最小程度的细胞凋亡,对p53磷酸化没有显著影响。最后,我们研究了CYP1B1活性抑制剂2,4,3',5'-四甲氧基芪和荧蒽的作用,CYP1B1在二苯并[a,l]芘的代谢活化中起核心作用。这两种化合物均以剂量依赖方式抑制二苯并[a,l]芘诱导的细胞凋亡,表明其干扰了后者的代谢活化。目前的数据表明,在与卵圆细胞具有共同表型特性的模型细胞系中,PAHs可被有效代谢形成最终的基因毒性代谢产物。肝祖细胞可能因此易受这种类型的基因毒性损伤,这使得WB - F344细胞系成为研究肝细胞中有机污染物基因毒性作用的有用工具。我们的结果还表明,与成熟肝细胞不同,CYP1B1可能是肝祖细胞中负责形成DNA加合物的主要酶。

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