Wise Laboratory of Environmental and Genetic Toxicology, Department of Pharmacology and Toxicology, University of Louisville, 500 S Preston St, Rm 1422, Louisville, KY, United States.
University of Southern Maine, Portland, ME, USA.
J Trace Elem Med Biol. 2020 Dec;62:126562. doi: 10.1016/j.jtemb.2020.126562. Epub 2020 May 26.
Hexavalent chromium [Cr(VI)] is a human lung carcinogen and global marine pollutant. High Cr concentrations, resembling the ones observed in occupationally exposed workers, have been observed in fin whales (Balaenoptera physalus) in the Gulf of Maine. This outcome suggests Cr might be disrupting the health of fin whale populations. Indeed, Cr in acute (24 h) exposure does cause toxicity in fin whale cells. However, human cell culture data indicate prolonged exposures (120 h) induce a higher amount of toxicity compared to 24 h exposure due to an inhibition of homologous recombination repair. However, whether prolonged exposure causes similar outcomes in fin whale cells is unknown.
Due to the importance of assessing prolonged exposure toxicity, this study focuses on characterizing acute and prolonged exposure of Cr(VI) in male and female fin whale cells.
Cytotoxicity was measured by the clonogenic assay, also known as colony forming assay, which measures the ability of cells to proliferate and form colonies after the treatment. DNA double strand breaks were analyzed by neutral comet assay. Clastogenicity was measured using the chromosome aberration assay. Intracellular Cr levels were measured with Graphite Furnace Atomic Absorption Spectrometry (GFAAS) with Syngistix Software.
In this study, we demonstrate that particulate Cr(VI) induces cytotoxicity and genotoxicity in a treatment-dependent manner after 24 h and 120 h exposures. Cytotoxicity levels were generally low with relative survival above 64 %. DNA double strand break data and chromosome aberration data were elevated after a 24 h exposure, but decreased after a 120 h exposure. While cytotoxicity was similar after 24 h and 120 h exposures, less DNA double strand breaks and chromosomal instability occurred with prolonged exposure.
Particulate Cr(VI) is cytotoxic and genotoxic to fin whale cells after acute and prolonged exposures. The reduction of genotoxicity we have observed after 120 h exposure may be partly explained by lower intracellular Cr levels after 120 h. However, the decrease in intracellular levels is not reflected by a similar decrease in chromosome aberrations suggesting other mechanisms may be at play. Male fin whale cells appear to be more susceptible to the genotoxic effects of particulate Cr(VI) while female cells are less susceptible possibly due to increased cell death of damaged cells, but more work is needed to clarify if this outcome reflects a sex difference or interindividual variability. Overall, the study shows particulate Cr(VI) does induce toxicity at both acute and prolonged exposures in fin whales cells indicating Cr(VI) exposure is a health risk for this species.
六价铬(Cr(VI))是一种人类肺部致癌物和全球海洋污染物。在缅因湾的长须鲸(Balaenoptera physalus)中,已经观察到类似于职业暴露工人的高浓度 Cr。这一结果表明 Cr 可能正在破坏长须鲸种群的健康。事实上,Cr 在急性(24 小时)暴露下确实会导致长须鲸细胞毒性。然而,人类细胞培养数据表明,由于同源重组修复的抑制,与 24 小时暴露相比,延长暴露(120 小时)会导致更高水平的毒性。然而,延长暴露是否会在长须鲸细胞中产生类似的结果尚不清楚。
由于评估延长暴露毒性的重要性,本研究侧重于描述雄性和雌性长须鲸细胞中 Cr(VI)的急性和延长暴露情况。
通过集落形成试验(也称为克隆形成试验)测量细胞毒性,该试验测量细胞在处理后增殖和形成集落的能力。通过中性彗星试验分析 DNA 双链断裂。使用染色体畸变试验测量断裂剂。使用 Graphite Furnace Atomic Absorption Spectrometry(GFAAS)和 Syngistix 软件测量细胞内 Cr 水平。
在这项研究中,我们证明了颗粒状 Cr(VI)在 24 小时和 120 小时暴露后以依赖于处理的方式诱导细胞毒性和遗传毒性。细胞存活率通常高于 64%,细胞毒性水平较低。24 小时暴露后 DNA 双链断裂数据和染色体畸变数据升高,但 120 小时暴露后降低。虽然 24 小时和 120 小时暴露后的细胞毒性相似,但延长暴露后发生的 DNA 双链断裂和染色体不稳定性较少。
颗粒状 Cr(VI)在急性和延长暴露后对长须鲸细胞具有细胞毒性和遗传毒性。我们在 120 小时暴露后观察到的遗传毒性降低可能部分归因于 120 小时后细胞内 Cr 水平降低。然而,细胞内水平的降低并没有反映出染色体畸变的类似减少,这表明可能存在其他机制在起作用。雄性长须鲸细胞似乎对颗粒状 Cr(VI)的遗传毒性更敏感,而雌性细胞则不那么敏感,这可能是由于受损细胞的细胞死亡增加,但还需要更多的工作来澄清这种结果是否反映了性别差异或个体间的变异性。总的来说,该研究表明颗粒状 Cr(VI)在长须鲸细胞中既会引起急性暴露毒性,也会引起延长暴露毒性,这表明 Cr(VI)暴露对该物种构成健康风险。