Citrinin Provoke DNA Damage and Cell-Cycle Arrest Related to Chk2 and FANCD2 Checkpoint Proteins in Hepatocellular and Adenocarcinoma Cell Lines

IF 3.9 3区医学 Q2 FOOD SCIENCE & TECHNOLOGY

Toxins Pub Date : 2024-07-17 DOI:10.3390/toxins16070321

Darija Stupin Polančec, Sonja Homar, Daniela Jakšić, Nevenka Kopjar, Maja Šegvić Klarić, Sanja Dabelić

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引用次数: 0

Abstract

Citrinin (CIT), a polyketide mycotoxin produced by Penicillium, Aspergillus, and Monascus species, is a contaminant that has been found in various food commodities and was also detected in house dust. Several studies showed that CIT can impair the kidney, liver, heart, immune, and reproductive systems in animals by mechanisms so far not completely elucidated. In this study, we investigated the CIT mode of action on two human tumor cell lines, HepG2 (hepatocellular carcinoma) and A549 (lung adenocarcinoma). Cytotoxic concentrations were determined using an MTT proliferation assay. The genotoxic effect of sub-IC50 concentrations was investigated using the alkaline comet assay and the impact on the cell cycle using flow cytometry. Additionally, the CIT effect on the total amount and phosphorylation of two cell-cycle-checkpoint proteins, the serine/threonine kinase Chk2 and Fanconi anemia (FA) group D2 (FANCD2), was determined by the cell-based ELISA. The data were analyzed using GraphPad Prism statistical software. The CIT IC50 for HepG2 was 107.3 µM, and for A549, it was >250 µM. The results showed that sensitivity to CIT is cell-type dependent and that CIT in sub-IC50 and near IC50 induces significant DNA damage and cell-cycle arrest in the G2/M phase, which is related to the increase in total and phosphorylated Chk2 and FANCD2 checkpoint proteins in HepG2 and A549 cells.

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橘皮苷在肝细胞癌和腺癌细胞株中引发与 Chk2 和 FANCD2 检查点蛋白相关的 DNA 损伤和细胞周期停滞

橘皮霉素（CIT）是一种由青霉、曲霉和大马士革霉产生的多酮类霉菌毒素，是一种在多种食品中发现的污染物，在室内灰尘中也有检出。多项研究表明，CIT 可损害动物的肾、肝、心脏、免疫和生殖系统，其作用机制至今尚未完全阐明。在本研究中，我们研究了 CIT 对两种人类肿瘤细胞系 HepG2（肝细胞癌）和 A549（肺腺癌）的作用模式。细胞毒性浓度是通过 MTT 增殖试验确定的。使用碱性彗星试验研究了 IC50 以下浓度的遗传毒性效应，并使用流式细胞仪研究了其对细胞周期的影响。此外，还利用细胞酶联免疫吸附法测定了 CIT 对两种细胞周期检查点蛋白（丝氨酸/苏氨酸激酶 Chk2 和范可尼贫血（FA）组 D2 (FANCD2)）的总量和磷酸化的影响。数据使用 GraphPad Prism 统计软件进行分析。CIT 对 HepG2 的 IC50 为 107.3 µM，对 A549 的 IC50 >250 µM。结果表明，对 CIT 的敏感性取决于细胞类型，IC50 以下和 IC50 附近的 CIT 会诱导显著的 DNA 损伤和细胞周期停滞在 G2/M 期，这与 HepG2 和 A549 细胞中 Chk2 和 FANCD2 检查点蛋白的总量和磷酸化量增加有关。

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来源期刊

Toxins TOXICOLOGY-

CiteScore

7.50

自引率

16.70%

发文量

765

审稿时长

16.24 days

期刊介绍： Toxins (ISSN 2072-6651) is an international, peer-reviewed open access journal which provides an advanced forum for studies related to toxins and toxinology. It publishes reviews, regular research papers and short communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced.