Conditionally reprogrammed human normal bronchial epithelial cells express comparable levels of cytochromes p450 and are sensitive to BaP induction
Conditionally reprogrammed human normal bronchial epithelial cells express comparable levels of cytochromes p450 and are sensitive to BaP induction
复制标题
条件性重编程的人正常支气管上皮细胞表达相当水平的细胞色素 p450，并且对 BaP 诱导敏感
DOI：
10.1016/j.bbrc.2018.08.001
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发表时间：
2018
影响因子：
3.1
通讯作者：
Li Hui
中科院分区：
文献类型：
--
作者：
Zhang Zhengbao;Bai Qing;Chen Yu;Ye Lina;Wu Xiaoting;Long Xiang;Ye Lin;Liu Jianjun;Li Hui
细胞色素P450(Cytochromes P450，Cyps)代谢数千种内源和外源化学物质，包括有毒化合物和药物。原代细胞寿命相对较短，在体外不能维持足够长时间的代谢酶CYPS的表达和活性。永生化的细胞系也不是进行毒性测试的理想细胞，因为它们的Cyps表达水平很低。在本研究中，我们使用条件重编程(CR)技术从三个人类供体(命名为CR-HNBE1-3)中建立了人正常支气管上皮细胞。这些CR细胞在规定的培养体系中可以在2个月内持续增殖超过50个PDs。在Matrigel 3D培养条件下，CR-HNBE细胞表现出正常的二倍体核型、对DNA损伤的正常反应和正常的分化潜能。CR-HNBE细胞表达基底上皮标记物细胞角蛋白14(CK14)和上皮分泌标记物粘蛋白5AC。最重要的是，CR-HNBE细胞表达的CYP1B1和CYP2E1水平与肺组织相当。与肺组织相比，这些CR细胞也表达类似的CYP1A1/CYP1A2、CYP2B6/CYP2C9/CYP2D6和CYP3A4/CYP3A5的mRNA。与毒理学领域广泛应用的永生化细胞系16HBE相比，这些CR细胞中的CYP1A1/CYP1B1的基础活性提高了3-6倍。我们的研究结果还表明，苯并(A)芘(BaP)可诱导CR-HNBE细胞中细胞色素P1A1和细胞色素P1A2的基因表达增加100倍。在CR-HNBE细胞中，BaP可诱导细胞色素P1A1/细胞色素P1B1的活性增加7~8倍，而在16HBE细胞中，可使细胞色素P1A1/细胞色素P1B1的活性增加2.5倍。综上所述，CR-HNBE细胞表达相当水平的细胞色素P450，并且对BaP的诱导敏感，将为体外毒性测试提供一个敏感的、生理的和有价值的模型。这是第一个关于正常人类呼吸道细胞可以长时间繁殖并保持相当水平的细胞周期蛋白的报道。
Cytochromes p450 (CYPs) metabolize thousands of endogenous and exogenous chemicals, including toxic compounds and drugs. The primary cells have relative short life span and are not able to sustain levels of metabolic enzymes CYPs expression and activity long enough in vitro. The immortalized cell lines are also not ideal for toxicity testing because of their low levels of CYPs expression. In this study, we established human normal bronchial epithelial cells using conditional reprogramming (CR) technique from three human donors (named as CR-HNBE1-3). These CR cells can proliferate continuously in defined culture system over 50 PDs within 2 months. The CR-HNBE cells exhibited the normal diploid karyotype, normal response to DNA damage and normal differentiation potential under the matrigel 3D culture condition. The CR-HNBE cells express the basal epithelial marker cytokeratin 14 (CK14) and epithelial secretory marker Mucin 5AC. Most importantly, CR-HNBE cells express comparable levels of CYP1B1 and CYP2E1 as those in lung tissue. These CR cells also express comparable mRNA of CYP1A1/CYP1A2, CYP2B6/CYP2C9/CYP2D6 and CYP3A4/CYP3A5 compared to the lung tissue. The basal activity of CYP1A1/CYP1B1 in these CR cells was 3–6 folds higher than that of 16HBE cells (an immortalized cell line widely used in toxicology field). Our data also demonstrated that Benzo(a)pyrene (BaP) induced up to 100 folds of mRNA expression of CYP1A1 or CYP1A2 in CR-HNBE cells. The activity of CYP1A1/CYP1B1 was induced by BaP up to 7–8 folds in CR-HNBE cells, while the activity of CYP1A1/CYP1B1 was induced maximum 2.5 folds in 16HBE cells. Taken together, CR-HNBE cells express comparable levels of CYPs and are sensitive to BaP induction, and will serve a sensitive, physiological and valuable in vitro toxicity testing model. This is the first report that normal human airway cells can be propagated for a long time and maintain comparable levels of CYPs.