Expression of multiple cytochrome P450 enzymes and multidrug resistance-associated transport proteins in human skin keratinocytes
Expression of multiple cytochrome P450 enzymes and multidrug resistance-associated transport proteins in human skin keratinocytes
复制标题
DOI：
10.1046/j.1523-1747.2001.01298.x
复制
发表时间：
2001-04-01
影响因子：
6.5
通讯作者：
Jugert, FK
中科院分区：
文献类型：
--
作者：
Baron, JM;Höller, D;Jugert, FK
细胞色素P450酶代谢各种内源性和外源性小分子量化合物。转运相关蛋白，如p -糖蛋白、多药耐药相关蛋白和肺耐药蛋白在耐药细胞系以及各种组织学来源的人类肿瘤（包括恶性黑色素瘤）中过度表达。对细胞色素酶和多药耐药相关转运蛋白在人类皮肤中的表达和功能知之甚少；因此，本研究的目的是分析细胞色素酶和多药耐药相关转运蛋白在组成条件下和不同诱导剂诱导后增殖的人表皮角质形成细胞中的表达模式。逆转录聚合酶链反应显示，角质形成细胞中细胞色素1A1、1B1、2B6、2E1和3A5组成性表达，地塞米松培养后细胞色素3A4表达，苯蒽诱导后细胞色素1A1在mRNA水平上表达增强。多药耐药相关转运蛋白逆转录-聚合酶链反应分析显示，多药耐药相关蛋白1、3-6和肺耐药蛋白在人上皮角质形成细胞中组成性表达，多药耐药1、2均为阴性，地塞米松诱导后1表达；免疫印迹法证实逆转录聚合酶链反应结果，发现地塞米松诱导后细胞色素1A1、2B6、2E1、3A、多药耐药相关蛋白1、3、5及多药耐药1均有表达。皮肤标本免疫组织显示细胞色素1A1、2B6、2E1和3A及多药耐药相关蛋白1和多药耐药相关蛋白1的免疫荧光阳性，催化法测定细胞色素1A1、2B、2E1和3A酶的组成活性，结果表明人皮肤角质形成细胞表达多种运输相关酶和解毒代谢酶。先前的研究表明，细胞色素酶和转运相关蛋白在药物的生物转化（I期）和抗转运（III期）处置中起互补作用，并作为药物生物利用度屏障协同作用。
Cytochrome P450 enzymes metabolize various endogenous and exogenous small molecular weight compounds. Transport-associated proteins, such as P-glycoprotein, multidrug resistance-associated protein and lung resistance protein are overexpressed in drug-resistant cell lines, as well as in human tumors from various histologic origins, including malignant melanoma, Little is known about the expression and function of cytochrome enzymes and multidrug resistance-associated transport proteins in human skin; therefore, the aim of this study was to analyze the expression pattern of cytochrome enzymes and multidrug resistance-associated transport proteins in proliferating human epidermal keratinocytes under constitutive conditions and after induction with various inducers. Reverse transcription-polymerase chain reaction revealed constitutive expression of cytochromes 1A1, 1B1, 2B6, 2E1, and 3A5 in keratinocytes and showed expression of cytochrome 3A4 after incubation with dexamethasone, The expression of cytochrome 1A1 was enhanced on the mRNA level after induction with benzanthracene. Reverse transcription-polymerase chain reaction analysis of the multidrug resistance-associated transport proteins revealed constitutive expression of multidrug resistance-associated proteins 1 and 3-6, and lung resistance protein in human epithelial keratinocytes and was negative for multidrug resistance 1 and 2, Expression of 1 was seen after induction with dexamethasone, Reverse transcription-polymerase chain reaction results were confirmed by immunoblots which showed expression of cytochromes 1A1, 2B6, 2E1, and 3A, multidrug resistance-associated proteins 1, 3, and 5 as well as multidrug resistance 1 after induction with dexamethasone. Immunohistology showed positive immunofluorescence in skin specimens for cytochromes 1A1, 2B6, 2E1, and 3A and multidrug resistance-associated protein 1 and multidrug resistance 1, Constitutive activity of cytochrome 1A1, 2B, 2E1, and 3A enzymes was measured by catalytic assays, These results show that keratinocytes of the human skin express various transport-associated enzymes and detoxifying metabolic enzymes. Previous studies have revealed that cytochrome enzymes and transport-associated proteins play complementary parts in drug disposition by biotransformation (phase I) and anti-transport (phase III) and act synergistically as a drug bioavailability barrier.