Research Article
In Vivo and In Vitro Genotoxic and Epigenetic Effects of Two
Types of Cola Beverages and Caffeine: A Multiassay Approach
Marcos Mateo-Fernández,
1
Tania Merinas-Amo,
1
Miguel Moreno-Millán,
1
Ángeles Alonso-Moraga,
1
and Sebastián Demyda-Peyrás
2
1
Department of Genetics, University of C´ ordoba, Rabanales Campus, 14071 C´ ordoba, Spain
2
Institute of Veterinary Genetics (IGEVET), Facultad de Ciencias Veterinarias, UNLP-CONICET, Universidad Nacional de La Plata,
1900 La Plata, Argentina
Correspondence should be addressed to Marcos Mateo-Fern´ andez; b42matfm@uco.es
Received 11 March 2016; Revised 14 May 2016; Accepted 5 June 2016
Academic Editor: Pravin K. Naoghare
Copyright © 2016 Marcos Mateo-Fern´ andez et al. Tis is an open access article distributed under the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Te aim of this work was to assess the biological and food safety of two diferent beverages: Classic Coca Cola6 (CCC) and Cafeine-
Free Coca Cola (CFCC). To this end, we determined the genotoxicological and biological efects of diferent doses of lyophilised
CCC and CFCC and Cafeine (CAF), the main distinctive constituent. Teir toxic/antitoxic, genotoxic/antigenotoxic, and chronic
toxicity (lifespan assay) efects were determined in vivo using the Drosophila model. Teir cytotoxic activities were determined
using the HL-60 in vitro cancer model. In addition, clastogenic DNA toxicity was measured using internucleosomal fragmentation
and SCGE assays. Teir epigenetic efects were assessed on the HL-60 methylation status using some repetitive elements. Te
experimental results showed a slight chemopreventive efect of the two cola beverages against HL-60 leukaemia cells, probably
mediated by nonapoptotic mechanisms. Finally, CCC and CAF induced a global genome hypomethylation evaluated in LINE-1
and Alu M1 repetitive elements. Overall, we demonstrated for the frst time the safety of this famous beverage in in vivo and in vitro
models.
1. Introduction
Diet may modify cancer risk and tumor behavior since
nongenotoxicological modulation as epigenetic regulatory
processes may be susceptible to changes caused by environ-
mental factors. Terefore, constituents in food and dietary
supplements could be involved in changes in the gene expres-
sion, increasing the risk of developing some type of cancer
all over the life inducing epigenetic changes [1, 2]. Geno-
toxicological screening tests have been extensively used over
time for assessing the health properties of compounds prior
to being considered as safe substances. Nowadays, the list of
foods with documented health-beneft activities is endless,
and scientifc evidence supporting the concept of health-
promoting food ingredients is steadily growing [3].
Originally developed as medical supplements, cola-based
drinks and several beverages such as beer and wine were
proposed as medicinal substances [4, 5]. However, a rela-
tionship between the consumption of these beverages and an
increase in the prevalence of several diseases such as child
obesity, diabetes, hypertension, and dental diseases was also
demonstrated [6–8]. In spite of the worldwide importance
and spread of cola beverages, studies assessing their efects
on health and wellbeing are quite scarce [9]. On the contrary,
cafeine (CAF), which is a key ingredient in cola beverages as
well as in cofee, tea, and some medicines, is one of the most
investigated substances, probably due to the lack of consistent
results over time [10–12]. In D. melanogaster, CAF has been
related to a positive lifespan increase [13], but the results were
contradictory when apoptotic and DNA-programmed frag-
mentation efects were studied [14, 15].
Drosophila is being used more frequently as a model for
many human diseases, including cancer [16–18]. Reiter et al.
Hindawi Publishing Corporation
BioMed Research International
Volume 2016, Article ID 7574843, 15 pages
http://dx.doi.org/10.1155/2016/7574843