Comparative study of Domoic Acid and Okadaic Acid induced-chromosomal abnormalities in the Caco-2 cell line.
Comparative study of Domoic Acid and Okadaic Acid induced-chromosomal abnormalities in the Caco-2 cell line.
复制标题
DOI：
10.3390/ijerph2006030001
复制
发表时间：
2006-03
影响因子：
--
通讯作者：
Creppy EE
中科院分区：
文献类型：
--
作者：
Carvalho PS;Catian R;Moukha S;Matias WG;Creppy EE
冈田酸（OA）是一种主要的贝类毒素，被认为是一种肿瘤促进剂，似乎与消化道癌症的发生有关。软骨藻酸（DA）是一种主要的贝类毒素，其遗传毒性和致癌性知之甚少。OA和DA都存在于海产品中，并且是人类健康问题。微核（MN）是由有丝分裂过程中由于纺锤体的失败或复杂的染色体构型造成的核分裂异常引起的，这些染色体构型在后期会造成问题。为了评价冈田酸（OA）和软骨藻酸（DA）诱导DNA损伤的能力，我们使用Caco-2细胞系进行了微核试验。为了区分OA和DA的致染色体断裂或非整倍体效应，采用细胞松弛素B结合Giemsa染色和/或吖啶橙子染色，通过分裂阻滞微核试验进行微核试验，同时采用浓缩的人泛着丝粒染色体涂料探针进行荧光原位杂交（FISH）。我们的结果表明，OA和DA显着增加MN在Caco-2细胞的频率。OA引起的微核见于单核细胞和双核细胞，而DA引起的微核主要见于双核细胞。荧光原位杂交结果表明，OA可诱导小鼠骨髓细胞产生着丝粒阳性微核，DA可增加无着丝粒信号的微核率。总之，OA和DA均具有致突变潜力，如在Caco-2细胞中通过诱导MN形成所揭示的。此外，OA诱导的整个染色体丢失表明一个特定的非整倍体潜力，而DA似乎只是致染色体断裂。目前，如果在体内达到所研究的浓度，则不能排除肠细胞可能受到DNA损伤的可能性，因为如果经常食用受污染的贝类，毒素浓度可能会低于监管限度。
Okadaic Acid (OA) the major diarrheic shellfish poisoning (DSP) toxin is known as a tumor promoter and seems likely implicated in the genesis of digestive cancer. Little is known regarding genotoxicity and carcinogenicity of Domoic Acid (DA), the major Amnesic Shellfish Poisoning (ASP) toxin. Both OA and DA occur in seafood and are of human health concerns. Micronuclei (MN) arise from abnormalities in nuclear division during mitosis due to a failure of the mitotic spindle or by complex chromosomal configurations that pose problems during anaphase. In order to evaluate the ability of okadaic acid (OA) and domoic acid (DA) to induce DNA damage we performed the micronucleus assay using the Caco-2 cell line. To discriminate between a clastogenic or aneugenic effect of OA and DA, the micronucleus assay was conducted by cytokinesis-block micronucleus assay using cytochalasin B with Giemsa staining and/or acridine orange staining, in parallel to fluorescence in situ hybridization (FISH) using a concentrated human pan-centromeric chromosome paint probe. Our results showed that OA and DA significantly increased the frequency of MN in Caco-2 cells. The MN caused by OA are found in mononucleated cells and binucleated cells, whereas those caused by DA are mainly in binucleated cells. The results of FISH analysis showed that OA induced centromere-positive micronuclei and DA increased the percentage of MN without a centromeric signal. In conclusion, both OA and DA bear mutagenic potential as revealed in Caco-2 cells by induction of MN formation. Moreover, OA induced whole chromosome loss suggesting a specific aneugenic potential, whereas DA seems simply clastogenic. At present, one cannot rule out possible DNA damage of intestinal cells if concentrations studied are reached in vivo, since this may happen with concentrations of toxins just below regulatory limits in case of frequent consumption of contaminated shell fishes.