title: Individual and combined effects of Aflatoxin B1, Deoxynivalenol and Zearalenone on HepG2 and RAW 264.7 cell lines.
abstract: To understand the combinatorial toxicity of mycotoxins, we measured the effects of individual, binary and tertiary combinations of Aflatoxin B1(AFB1), Deoxynivalenol (DON) and Zearalenone (ZEN) on the cell viability and cellular perturbations of HepG2 and RAW 264.7 cells. The nature of mycotoxins interactions was assessed using mathematical modeling (Chou-Talalay). Mechanisms of cytotoxicity were studied using high content screening (HCS) that probed cytotoxicity responses, such as changes in intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP), intracellular calcium ([Ca2+]i) flux, and cell membrane damage. Our results showed that individual cytotoxicity of mycotoxins in a decreasing order was DON>AFB1>ZEN. Varying combinations of mycotoxins at differing concentrations showed different types of interactions. Most of the mixtures showed increasing toxic effects-synergism and/or addition while antagonistic effects were observed with combination of AFB1+ZEN. Generally, combination of mycotoxins showed significantly increased intracellular ROS production and [Ca2+]iflux, and decreased MMP in both cell lines, showing that the synergistic and additive effects of mycotoxin combination originate from perturbations of multiple cellular functions. Additionally, this study demonstrated the applicability of HCS for gaining mechanistic understanding on the toxicity of individual as well as combinatorial mycotoxins, and also provided scientific bases for formulating regulatory policies.
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Food Safety and Quality Control of Jiangsu Province, Jiangnan University, Wuxi 214122, PR China.
authors.forename: Hongyuan
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Centre for Sustainable Nanotechnology, School of Chemical & Life Sciences, Nanyang Polytechnic, Singapore 569830, Singapore. Electronic address: saji.george@mcgill.ca.
authors.forename: Saji
authors.initials: S
authors.lastname: George
authors.affiliation: Centre for Sustainable Nanotechnology, School of Chemical & Life Sciences, Nanyang Polytechnic, Singapore 569830, Singapore.
authors.forename: Crystal
authors.initials: C
authors.lastname: Hay
authors.affiliation: Centre for Sustainable Nanotechnology, School of Chemical & Life Sciences, Nanyang Polytechnic, Singapore 569830, Singapore.
authors.forename: Joel
authors.initials: J
authors.lastname: Lee
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Food Safety and Quality Control of Jiangsu Province, Jiangnan University, Wuxi 214122, PR China.
authors.forename: He
authors.initials: H
authors.lastname: Qian
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Food Safety and Quality Control of Jiangsu Province, Jiangnan University, Wuxi 214122, PR China. Electronic address: sxlzzz@jiangnan.edu.cn.
authors.forename: Xiulan
authors.initials: X
authors.lastname: Sun
mesh_terms: D016604:Aflatoxin B1 / Q000008:administration & dosage / Q000633:toxicity*; D000818:Animals; D004305:Dose-Response Relationship, Drug; D056945:Hep G2 Cells / Q000187:drug effects; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D009183:Mycotoxins / Q000008:administration & dosage / Q000633:toxicity; D000067996:RAW 264.7 Cells / Q000187:drug effects; D018675:Toxicity Tests / Q000379:methods*; D014255:Trichothecenes / Q000008:administration & dosage / Q000633:toxicity*; D015025:Zearalenone / Q000008:administration & dosage / Q000633:toxicity*
chemical_list: D009183:Mycotoxins; D014255:Trichothecenes; D015025:Zearalenone; D016604:Aflatoxin B1; C007262:deoxynivalenol
keywords: Chou-Talalay method; Combinatorial cytotoxicity; High content screening; Mammalian cell line; Mycotoxin