Matrix metalloproteinases and their inhibitors as biomarkers for metal toxicity in vitro
Matrix metalloproteinases and their inhibitors as biomarkers for metal toxicity in vitro
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DOI：
10.1016/j.tiv.2006.02.004
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发表时间：
2006-10-01
影响因子：
3.2
通讯作者：
Giuliano, Mariateresa
中科院分区：
文献类型：
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作者：
Cammarota, Marcella;Lamberti, Monica;Giuliano, Mariateresa
暴露于镍和铬及其化合物会对健康产生不良影响。这些金属是两种人类致癌物，其发病机制涉及主动细胞外基质降解和重塑。在这项工作中，我们比较了体外暴露于镍和铬的角质形成细胞系（HaCat）的影响。基质金属蛋白酶基因的调节被用作化学损伤的生物标志物。将融合细胞持续暴露于亚毒性浓度的铬和镍（10（-5）和10（-7）M）中长达72 h。提取总RNA，RT-PCR分析特异性基质金属蛋白酶及其抑制剂基因表达。流式细胞仪检测细胞周期的变化。镍和铬表现出不同的结果，在镍处理的细胞中MMP-2 mRNA的产生上调，而铬暴露下调MMP-2 mRNA的产生。这一结果可能与铬处理细胞中组织抑制因子-1（TIMP-1）mRNA的过早（6 h）过表达有关。细胞周期分析显示，4 N DNA的细胞增加。这些结果可以解释为通过TIMP-1的抗凋亡作用逃避金属诱导的凋亡的细胞的存活反应。这些包含遗传毒性损伤的细胞可能具有选择性增殖优势，因此代表了可能出现退化变体的前体库。为了研究化学损伤是否可逆，仅刺激亚融合细胞24小时，然后更换培养基而不含金属。细胞能够从镍暴露中恢复，相对于对照，仅显示出特异性mRNA表达和细胞周期改变的微弱改变。铬引起的损害是不可逆转的。我们的研究结果表明，金属毒性和MMPs及其抑制剂的表达之间存在关联。这些生物标志物可能是潜在的有用的阐述化学毒性的预测模型。(c)2006爱思唯尔有限公司保留所有权利。
Exposure to nickel and chromium, and their compounds, has been associated with adverse health effects. These metals are two human carcinogens whose pathogenesis involves active extracellular matrix degradation and remodelling. In this work we have compared the effects of in vitro exposure to nickel and chromium of a keratinocyte cell line (HaCat). The modulation of matrix metalloproteinase genes was used as biomarker of chemical damage. Confluent cells were constantly exposed to subtoxic chromium and nickel concentrations (10(-5) and 10(-7) M) up to 72 h. Total RNA was extracted and specific matrix metalloproteinase, and inhibitor, gene expression was analyzed by RT-PCR. Moreover, cell cycle alterations were evaluated by flow cytometry. Nickel and chromium showed different results, with an upregulation of MMP-2 mRNA production in nickel-treated cells while chromium exposure down-regulated MMP-2 mRNA production. This result could be correlated to the precocious (6 h) over-expression of tissue inhibitor-1 (TIMP-1) mRNA in chromium-treated cells. Cell cycle analysis showed and increase of cells with 4N DNA. These results could be explained as a survival response of cells that escape metal induced apoptosis through the anti-apoptotic effects of TIMP-1. These cells that encompass the genotoxic insult may have a selective proliferation advantage, and therefore represent the precursor pool from which degenerating variants may emerge. To study if the chemical damage was reversible, subconfluent cells were stimulated only for 24 h, then the medium was replaced without metal. Cells were able to recover from nickel exposure, showing only weak alterations in specific mRNA expression and cell cycle alteration respect to control. Chromium-induced damage was irreversible. Our results demonstrated that there is an association between metal toxicity and expression of MMPs and their inhibitors. These biomarkers could be potentially useful to elaborate a prediction model of chemical toxicity. (c) 2006 Elsevier Ltd. All rights reserved.