title: In silico studies with human DNA topoisomerase-II alpha to unravel the mechanism of in vitro genotoxicity of benzene and its metabolites.
abstract: Exposure of humans to benzene present in environment may lead to adverse chronic effects-even at the genetic level. However, the mechanism of its genotoxicity is not well understood. In the present study, in vitro genotoxicity of benzene (BZ) and its major metabolites [p-benzoquinone (BQ), hydroquinone (HQ), catechol (CT), 1,2,4-benzenetriol (BT) and trans-trans muconic acid (MA)] at concentrations 0.5-50 microM, was assessed in Chinese hamster ovary (CHO) cells employing the alkaline Comet assay, cytokinesis blocked micronucleus (CBMN) assay, flow cytometric analysis of micronucleus (flow MN) and chromosome aberration (CA) test. The data revealed significant (P<0.05) concentration-dependent response in all end points. HQ was found to be the most potent DNA damaging metabolite in the Comet assay followed by BQ>BT>CT>BZ>MA. Both CBMN and flow MN assays revealed a good correlation in their results, where BQ and MA exhibited maximum and minimum micronucleus induction respectively. Significant chromosomal aberrations were induced mainly by BQ, BT and HQ, with moderate response shown by CT and BZ and least by MA. The results demonstrated the utility of sensitive techniques like Comet assay and flow cytometry for determination of MN, to quantify in vitro genotoxicity at low levels and also suggested that partly non-repaired DNA damage could cause adverse health effects in human population exposed to benzene. In silico studies using different endpoints of genotoxicity and molecular docking studies with human topoisomerase-II alpha, a major DNA repair enzyme were also conducted. These corroborated the results obtained from the in vitro data, pointing to a direct relationship of the observed genotoxicity with the structural properties and various interactions of metabolites with the enzyme. This comprehensive study demonstrated that genotoxicity of benzene in mammalian cells is mainly due to the inhibition of topoisomerase by the metabolites.
authors.affiliation: Developmental Toxicology Division, Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226 001, India. pandeyalok2002@yahoo.co.in
authors.forename: Alok Kumar
authors.initials: AK
authors.lastname: Pandey
authors.forename: Deepak
authors.initials: D
authors.lastname: Gurbani
authors.forename: Mahima
authors.initials: M
authors.lastname: Bajpayee
authors.forename: Devendra
authors.initials: D
authors.lastname: Parmar
authors.forename: Subhash
authors.initials: S
authors.lastname: Ajmani
authors.forename: Alok
authors.initials: A
authors.lastname: Dhawan
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000378:metabolism*; D001554:Benzene / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D001555:Benzene Derivatives / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D016466:CHO Cells; D020134:Catalytic Domain; D002869:Chromosome Aberrations; D020552:Comet Assay; D003198:Computer Simulation; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D015162:Micronucleus Tests; D008954:Models, Biological; D008958:Models, Molecular; D009153:Mutagens / Q000378:metabolism / Q000633:toxicity*; D021281:Quantitative Structure-Activity Relationship
chemical_list: D000951:Antigens, Neoplasm; D001555:Benzene Derivatives; D004268:DNA-Binding Proteins; D009153:Mutagens; D004250:DNA Topoisomerases, Type II; D001554:Benzene