Effects of Nicotine, Rifampicin and Benzo[a]pyrene on the Expression of Cytochrome P450 in Human Hepatocellular Carcinoma Cells
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Abstract
CYP1 and CYP2, members of cytochrome P450 enzymes (CYP), are mainly expressed in liver and responsible for several chemicals in daily life. Nicotine, a compound in cigarettes, is metabolized by CYP2A6 and CYP2A13. Benzo[a]pyrene (BaP), a carcinogen is found in grilled food and smoke from engines, is metabolized by CYP1A2. BaP is also an inducer of several CYP isoforms including CYP1A2. Rifampicin, an anti-tuberculosis drug, has potency to induce several CYPs and causes clinical drug interactions. The objective of this study was to determine the effect of nicotine, rifampicin, and BaP on the expression of CYP1A2, CYP2A6, CYP2A13, and CYP2E1 in human hepatocellular carcinoma (HepG2) cells. Methods: HepG2 was treated with nicotine (1, 10, and 100 µM), rifampicin (10 µM), BaP (10 µM), and 0.1% DMSO (as the control) for 24 h before total RNA extraction. After that, the expression of CYP1A2, CYP2A6, CYP2A13, CYP2E1, and AhR mRNA was determined using RT-qPCR, compared to a reference gene, GAPDH. Results: CYP1A2 was induced by low dose of nicotine (1 µM), same as BaP. Moreover, nicotine and BaP can induce the expression of CYP2A6 and CYP2A13 in dose-dependent pattern. CYP2E1 was induced by high dose of nicotine (100 µM), same as BaP. Rifampicin significantly induced the expression of CYP2A13 and CYP2E1. Interestingly, only nicotine at the low dose induced arylhydrocarbon receptor (AhR). Conclusion: Nicotine induced the expression of CYP2A6, CYP2A13, CYP1A2, and CYP2E1 mRNAs, comparable to BaP, and the regulatory pathway of CYPs by nicotine might related to AhR, at least in part.
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