Induction and regulation of xenobiotic-metabolizing cytochrome P450s in the human A549 lung adenocarcinoma cell line
Induction and regulation of xenobiotic-metabolizing cytochrome P450s in the human A549 lung adenocarcinoma cell line
复制标题
DOI：
10.1165/ajrcmb.22.3.3845
复制
发表时间：
2000-03-01
影响因子：
6.4
通讯作者：
Raunio, H
中科院分区：
文献类型：
--
作者：
Hukkanen, J;Lassila, A;Raunio, H
几种细胞色素P450(CYP)酶在人肺中表达，参与代谢失活和许多外源性和内源性化合物的激活。在这项研究中，利用定性逆转录/聚合酶链式反应(RT-PCR)研究了所有已知的外源代谢CYP基因在人肺泡II型细胞来源的A549腺癌细胞系中的表达模式。此外，用定量RT-PCR方法研究了化学物质对细胞色素P450 1和细胞色素P3A亚家族成员的诱导机制。在A549细胞中检测到CYP1A1、1B1、2B6、2C、2E1、3A5和3A7的信使RNA(MRNAs)的表达。细胞色素P1A2、2A6、2A7、2A13、2F1、3A4和4B1的mRNA量均低于检测限。2，3，7，8-四氯二苯并对二恶英(TCDD)可诱导细胞色素P1A1和细胞色素P1B1mRNAs分别增加56倍和2.5倍。地塞米松诱导的细胞色素P3A5为8倍，苯巴比妥诱导为11倍。使用的任何一种典型的细胞色素P3A4诱导剂都不能诱导细胞色素P3A4。酪氨酸激酶抑制剂金雀异黄素和蛋白激酶C抑制剂星形孢子素可阻断TCDD诱导的细胞色素P1A1的诱导，但不影响细胞色素P1B1的诱导。蛋白磷酸酶抑制剂冈田酸和花盏花素A对细胞色素P1B1的TCDD诱导有轻微的促进作用，但对细胞色素P1A1的诱导作用不明显。这些结果表明，在人肺上皮细胞中，细胞色素P4501A1和细胞色素P1B1B1RUE的表达存在差异，首次提出糖皮质激素对人肺上皮细胞中细胞色素P3A5的诱导作用。这些结果表明，A549细胞系保留了人肺上皮细胞CYP表达的一些特征，是一种有价值的诱导肺CYP系统的机制研究模型。
Several cytochrome P450 (CYP) enzymes are expressed in the human lung, where they participate in metabolic inactivation and activation of numerous exogenous and endogenous compounds. In this study, the expression pattern of all known xenobiotic-metabolizing CYP genes was characterized in the human alveolar type II cell-derived A549 adenocarcinoma cell line using qualitative reverse transcriptase/polymerase chain reaction (RT-PCR). In addition, the mechanisms of induction by chemicals of members in the CYP1 and CYP3A subfamilies were assessed by quantitative RT-PCR. The expression of messenger RNAs (mRNAs) of CYPs 1A1, 1B1, 2B6, 2C, 2E1, 3A5, and 3A7 was detected in the A549 cells. The amounts of mRNAs of CYPs 1A2, 2A6, 2A7, 2A13, 2F1, 3A4, and 4B1 were below the limit of detection. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced CYP1A1 and CYP1B1 mRNAs 56-fold and 2.5-fold, respectively. CYP3A5 was induced 8-fold by dexamethasone and 11-fold by phenobarbital. CYP3A4 was not induced by any of the typical CYP3A4 inducers used. The tyrosine kinase inhibitor genistein and the protein kinase C inhibitor staurosporine blocked TCDD-elicited induction of CYP1A1, but they did not affect CYP1B1 induction. Protein phosphatase inhibitors okadaic acid and calyculin A enhanced TCDD-induction of CYP1B1 slightly, but had negligible effects on CYP1A1 induction. These results suggest that CYP1A1 and CYP1B1 rue differentially regulated in human pulmonary epithelial cells and give the first indication of the induction of CYP3A5 by glucocorticoids in human lung cells. These results establish that having retained several characteristics of human lung epithelial cell CYP expression, the A549 lung cell line is a valuable model for mechanistic studies on induction of the pulmonary CYP system.