Mycotoxins’ Activity at Toxic and Sub-Toxic Concentrations: Differential Cytotoxic and Genotoxic Effects of Single and Combined Administration of Sterigmatocystin, Ochratoxin A and Citrinin on the Hepatocellular Cancer Cell Line Hep3B

Nikolia Αnninou,Μichail Pitiakoudis,Constantinos Simopoulos,Fotini Papachristou,Ekaterini Chatzaki

doi:10.3390/ijerph110201855

Nikolia Αnninou, Μichail Pitiakoudis + Show 3 more

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https://doi.org/10.3390/ijerph110201855

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Abstract

Food safety organizations indicate the likelihood of constant human and animal exposure to mycotoxin mixtures as a possible negative public health impact. Risk assessment demonstrates that certain mycotoxins of Aspergillus and Penicillium spp. are toxic and hold a significant genotoxic efficacy at nanomolar concentrations. The aim of the current study was to investigate the potential cytogenetic effects of sterigmatocystin (STER), ochratoxin A (OTA) and citrinin (CTN) alone or in combination, at pM to μΜ concentrations, on the human hepatocellular cancer cell line Hep3B. MTT reduction, mitotic divisions, cell cycle delays and sister chromatid exchange rates (SCE) were determined as endpoints of metabolic activity, cytotoxicity, cytostaticity, and genotoxicity, respectively. All mycotoxin treatments induce SCE rates from 10−12 M, while their cytotoxic and cytostatic potential varies. In PRI and MI assays, but not at MTT, STER alone or in combination with OTA + CTN appeared cytostatic and cytotoxic, even at 10−12 M, while CTN alone and all other combinations displayed substantial cellular survival inhibition in doses ≥ 10−8 M. Co-administration of STER + OTA or STER + CTN in concentrations ≤ 10−1 M, increased the MI and MTT activity, while it did not affect the PRI. Mycotoxin co-treatments revealed in general similar-to-additive or antagonistic genotoxic and cytotoxic effects. Our results for the first time describe that STER alone or in combination with OTA and/or CTN share a cytotoxic and cytogenetic potential even at picoMolar concentrations on human hepatoma cells in vitro.

Highlights

Mycotoxins are secondary metabolic products of molds, whereby the major mycotoxin-producing fungi are certain Aspergillus, Fusarium and Penicillium species [1]
STER (C) ochratoxin A (OTA) and CTN (E) STER and OTΑ for 24 and 48h; Bars represent mean values of every concentration at each time point, which was evaluated at three individual experiments, performing measurements at a number of 8-plicates each time (8-plicates, n = 3)
Evaluation of MTT activity in Hep3B cells after co-administration of (A) CTN, OTA and STER for 24 and 48h; Bars represent mean values of every concentration at each time point, which was evaluated at three individual experiments, performing measurements at a number of 8-plicates each time (8-plicates, n = 3)

Summary

Introduction

Mycotoxins are secondary metabolic products of molds, whereby the major mycotoxin-producing fungi are certain Aspergillus, Fusarium and Penicillium species [1]. Mycotoxins (in native and metabolized conjugated form) possess potentially toxic and even carcinogenic (after long-term exposure) properties [1,2,3,4,5,6,7]. Their impact on public health depends on the consumption of mycotoxin-contaminated processed agricultural and animal-derived products or meat [2,4]. According to in vivo as well in vitro studies, it was more cytotoxic than aflatoxin in human adenocarcinoma lung cells A549 and human esophageal epithelial cells Het-1A and exhibited a mutagenic potential [14]

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