Evaluation of Aflatoxin M1 Effects on the Metabolomic and Cytokinomic Profiling of a Hepatoblastoma Cell Line.

Silvia Marchese,Lorella Severino,Alfredo Budillon,Susan Costantini,Angela Sorice,Salvatore Florio,Andrea Ariano

doi:10.3390/toxins10110436

Silvia Marchese, Lorella Severino + Show 5 more

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https://doi.org/10.3390/toxins10110436

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Abstract

Hepatoblastoma incidence has been associated with different environmental factors even if no data are reported about a correlation between aflatoxin exposure and hepatoblastoma initiation. Considering that hepatoblastoma develops in infants and children and aflatoxin M1 (AFM1), the aflatoxin B1 (AFB1) hydroxylated metabolite, can be present in mothers’ milk and in marketed milk products, in this study we decided to test the effects of AFM1 on a hepatoblastoma cell line (HepG2). Firstly, we evaluated the effects of AFM1 on the cell viability, apoptosis, cell cycle, and metabolomic and cytokinomic profile of HepG2 cells after treatment. AFM1 induced: (1) a decrease of HepG2 cell viability, reaching IC50 at 9 µM; (2) the blocking of the cell cycle in the G0/G1 phase; (3) the decrease of formiate levels and incremented level of some amino acids and metabolites in HepG2 cells after treatment; and (4) the increase of the concentration of three pro-inflammatory cytokines, IL-6, IL-8, and TNF-α, and the decrease of the anti-inflammatory interleukin, IL-4. Our results show that AFM1 inhibited the growth of HepG2 cells, inducing both a modulation of the lipidic, glycolytic, and amino acid metabolism and an increase of the inflammatory status of these cells.

Highlights

Hepatoblastoma is a common hepatic cancer of infancy and childhood, which occurs mainly in the first years of life [1]
To assess the concentration at which cell growth was inhibited by 50%, we performed sulforhodamine B (SRB) assay on a HepG2 cell line treated with different concentrations of aflatoxin M1 (AFM1)
Considering that hepatoblastoma develops in infants and children, and AFM1 can be present in mothers milk and in marketed milk products, our aim was to evaluate if AFM1 affects cell growth, metabolism, and inflammatory status of HepG2, a hepatoblastoma cell line

Summary

Introduction

Hepatoblastoma is a common hepatic cancer of infancy and childhood, which occurs mainly in the first years of life [1]. Hepatoblastoma patients have elevated levels of α-fetoprotein (AFP), a useful marker of diagnosis and of treatment response [3]. It is reported that CTNNB1 mutations and the Wnt pathway, which regulates this gene, are involved in the initiation of this cancer. Knocked out CTNNB1 in a murine model, and demonstrated that partial hepatectomy induced hepatic cell proliferation and growth. This study evidenced that CTNNB1 abnormalities collectively account for most of the genetic defects in hepatoblastoma and its increased levels are present in almost all Toxins 2018, 10, 436; doi:10.3390/toxins10110436 www.mdpi.com/journal/toxins

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