Nicotine derived genotoxic effects in human primary parotid gland cells as assessed in vitro by comet assay, cytokinesis-block micronucleus test and chromosome aberrations test
Nicotine derived genotoxic effects in human primary parotid gland cells as assessed in vitro by comet assay, cytokinesis-block micronucleus test and chromosome aberrations test
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DOI：
10.1016/j.tiv.2014.03.012
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发表时间：
2014-08-01
影响因子：
3.2
通讯作者：
Kleinsasser, Norbert H.
中科院分区：
文献类型：
--
作者：
Ginzkey, Christian;Steussloff, Gudrun;Kleinsasser, Norbert H.
在包括唾液腺细胞在内的不同人类细胞中描述了尼古丁的遗传毒性作用。根据使用治疗性尼古丁斑块的吸烟者或患者唾液中的高尼古丁浓度，进行了当前的研究，以评估尼古丁在人唾液腺细胞中尼古丁的遗传毒性潜力。在没有外源代谢激活的情况下，在1 Mu M和1000 MU M之间含量1 h。腺泡表型通过α-淀粉酶的免疫荧光染色证明。使用彗星测定，微核测试和染色体畸变测试评估了遗传毒性作用。细胞毒性和凋亡是通过锥虫蓝色排除测试确定的，CAS-PASE-3测定法。NICOTINE能够在所有三种测定中诱导遗传毒性作用。染色体畸变测试是最敏感的，数值和结构性（染色单体型和染色体类型）的畸变在> = 1 Mu M处观察到，而在10 mu m和DNA损伤下，微核频率的增加，如在10 mu m和DNA损伤下的增加。在> 100 mu的情况下，彗星测定法被认为没有细胞毒性损伤或凋亡的影响。尼科宁作为唾液腺中肿瘤启动的可能危险因素仍在有争议上讨论。我们的结果表明，尼古丁在唾液腺细胞中诱导遗传毒性作用的潜力。这些结果是在与口服或经透皮暴露后发现的唾液尼古丁水平相似的，建议有必要仔细监测人类中尼古丁的使用。 （c）2014 Elsevier Ltd.保留所有权利。
Genotoxic effects of nicotine were described in different human cells including salivary gland cells. Based on the high nicotine concentration in saliva of smokers or patients using therapeutic nicotine patches, the current study was performed to evaluate the genotoxic potential of nicotine in human salivary gland cells.Therefore, primary salivary gland cells from 10 patients undergoing parotid gland surgery were exposed to nicotine concentrations between 1 mu M and 1000 mu M for 1 h in the absence of exogenous metabolic activation. The acinar phenotype was proven by immunofluorescent staining of alpha-amylase. Genotoxic effects were evaluated using the Comet assay, the micronucleus test and the chromosome aberration test. Cytotoxicity and apoptosis were determined by trypan blue exclusion test and Cas-pase-3 assay.Nicotine was able to induce genotoxic effects in all three assays. The chromosome aberration test was the most sensitive and increases in numerical and structural (chromatid-type and chromosome-type) aberrations were seen at >= 1 mu M, whereas increases in micronuclei frequency were detected at 10 mu M and DNA damage as measured in the Comet assay was noted at >100 mu M. No cytotoxic damage or influence of apoptosis could be demonstrated.Nicotine as a possible risk factor for tumor initiation in salivary glands is still discussed controversially. Our results demonstrated the potential of nicotine to induce genotoxic effects in salivary gland cells. These results were observed at saliva nicotine levels similar to those found after oral or transdermal exposure to nicotine and suggest the necessity of careful monitoring of the use of nicotine in humans. (C) 2014 Elsevier Ltd. All rights reserved.