Effects of chitosan-stabilized selenium nanoparticles on cell proliferation, apoptosis and cell cycle pattern in HepG2 cells: Comparison with other selenospecies

doi:10.1016/j.colsurfb.2014.06.062

Colloids and Surfaces B: Biointerfaces

Volume 122, 1 October 2014, Pages 184-193

https://doi.org/10.1016/j.colsurfb.2014.06.062 Get rights and content

Highlights

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A systematic comparison of the effect of Ch-SeNPs vs. other selenocompounds on hepatocarcinoma cells in terms of proliferation, apoptosis and cell cycle pattern has been carried out.
•
Cells exposed to Se(VI), seleno methylselenocysteine and selenomethionine do not exhibit a different behavior as compared to control cells at the tested concentrations.
•
Se(IV) induce a significant degree of apoptosis proving to be the most toxic selenospecies assayed.
•
Ch-SeNPs and selenocystine exposure induced a cell cycle arrest at the S-G2/M phase.
•
Ch-SeNPs showed a unique potential for targeting mitosis by partially inhibiting the expression of cyclin-dependent kinase 1 (Cdk1).

Abstract

Selenium is an essential element that plays an important role in many biological functions. Many studies have reported the potential beneficial effects of Se intake for cancer therapy and prevention, which are not only dose-dependent but also closely related to the properties of specific selenospecies. Selenium nanoparticles are considered a novel selenium compound with excellent antioxidant properties; however, little is known about the properties of selenium nanoparticles in comparison to other well-studied selenospecies. Here, we combined different independent bioanalytical approaches to carry out a comparison between the effects of selenium nanoparticles and other selenocompounds (inorganic and organic selenospecies) using an in-vitro model. The bioanalytical characterization of different parameters such as cell proliferation, apoptosis and cell cycle pattern on HepG2 cells has shown the unique properties of this relatively novel compound that support and complete prior evidences for future applications as chemotherapeutic agent.

Graphical abstract

Introduction

Selenium is an essential and unique micronutrient that plays an important role in regulating the functions of many intracellular proteins. Fairweather-Tait et al. [1] recently reviewed the relationship between selenium intake/status and several health outcomes such as cancer, cardiovascular disease, diabetes and male fertility. Low selenium status has been associated with increased risk of mortality, poor immune function, and cognitive decline [2].

The benefits of Se as a potential chemopreventive or chemotherapeutic agent have been extensively studied [3], [4], [5], [6]. Although the epidemiological data are somewhat controversial, it seems that both, inorganic and organic forms of Se, negatively affect cancer progression [7]. In addition, it is known that selenoproteins also play important roles in tumor development [8]. However, it is important to consider that the dose and the chemical form of selenium have a significant influence on these effects [9], [10], [11], [12], [13].

During the last years, attention on nanotechnology and nanomaterials has reached high relevance due to their multiple applications in several fields, including biomedical research [14]. Selenium nanoparticles (SeNPs) are considered a novel selenium compound with excellent antioxidant properties and lower toxicity than other selenospecies [15], [16]. Zhang et al. [17] demonstrated that SeNPs and Se(IV) had a similar bioavailability in rats and antioxidant effect in cells. On the other hand, SeNPs have been proved to present a better selectivity between normal and cancer cells than Se(IV) at similar concentrations [18]. Moreover, SeNPs have been appointed as a potential anticancer drug [19], [20] due to its ability to inhibit the growth of cancer cells through induction of S phase arrest [21]. In fact, the relationship between nanoparticles (not only in the case of SeNPs) and cell cycle life has been recently reviewed by Kim et al. [22] and Mahmoudi et al. [23].

In the present work, we compare the effect of chitosan-stabilized selenium nanoparticles (Ch-SeNPs) and other inorganic and organic selenospecies including Se(IV), Se(VI), selenomethionine (SeMet), selenocystine (SeCys₂) and seleno methylselenocysteine (Se-MeSeCys) on hepatocarcinoma (HepG2) cells in terms of viability, proliferation and migration. We also evaluate the capacity of the different species to induce apoptosis and/or cell cycle arrest. Finally, we consider the relevance of cyclin-dependent kinase 1 (Cdk1) in governing the mitotic cell death, which constitutes a current potential mechanism for cancer therapy. Thus, we study the inhibition degree of Cdk1 after exposing the cells to the above mentioned species in order to evaluate whether the properties exhibited by SeNPs are unique or shared by other selenocompunds.

Section snippets

Chemicals

Materials and chemicals for electrophoresis were from BioRad (Madrid, Spain). Sodium selenite, sodium selenate, selenomethionine, selenocystine, seleno methylselenocysteine, Dulbecco's modified Eagle's medium (DMEM), MTT reagent, Mowiol^® 4-88, chitosan, triton, paraformaldehyde (PFA), 4′,6-diamidino-2-phenylindole (DAPI) and Anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) were purchased from Sigma-Aldrich. Trypsin/EDTA and phosphate buffer saline (PBS) were purchased from Lonza (Basel,

Synthesis, characterization and stability of chitosan-stabilized selenium nanoparticles

It has been demonstrated that monosaccharides (glucose), oligosaccharides (sucrose) and polysaccharides (chitosan) modify the size, morphology and stability of SeNPs in liquid dispersions [24]. Previous studies have shown how encapsulation of selenite into chitosan significantly improve the antioxidant properties and promote a high retention of selenium in cells [25], [26]; thus, we used chitosan to enwrappe SeNPs by forming strong hydrogen bonds between the hydroxyl groups of chitosan and the

Conclusions

To the best of our knowledge, this is the first time that a systematic comparison of the effect of Ch-SeNPs with other inorganic and organic selenospecies on the behavior of cancer cells has been carried out. Our data, strongly suggest that cells exposed to Ch-SeNPs and SeCys₂ undergo similar alterations in terms of cell viability, proliferation, migration and cell cycle arrest at the S-G2/M phase. Cells exposed to Se(IV) showed evident signs of toxicity such as strong induction of apoptosis

Acknowledgements

Authors thank the Spanish Ministry of Economy and Competitiveness (grants CTQ2010-18644 and CTQ2011-28328C02-01), the Comunidad de Madrid (Spain) (Analisyc II) and the European FEDER programme (grants AS2009/AGR-1464, ANALISYC-II, Interreg European Project Orque-Sudoe). The authors thank Blanca González, Sandra Sánchez and Montserrat Colilla for their valuable help in synthesizing the Ch-SeNPs. The authors also thank Maria Luisa Garcia and Agustin Fernandez for electron microscopy services

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¹: These authors contributed equally to this work.

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Effects of chitosan-stabilized selenium nanoparticles on cell proliferation, apoptosis and cell cycle pattern in HepG2 cells: Comparison with other selenospecies

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Chemicals

Synthesis, characterization and stability of chitosan-stabilized selenium nanoparticles

Conclusions

Acknowledgements

Lancet

J. Trace Elem. Med. Biol.

Mutat. Res.

Trends Anal. Chem.

Free Radical Biol. Med.

Colloids Surf., B: Biointerfaces

Biomaterials

Colloids Surf., B: Biointerfaces

Mater. Lett.

Carbohydr. Polym.

J. Nutr. Biochem.

Toxicol. In Vitro

Curr. Opin. Genet. Dev.

Trends Biochem. Sci.

J. Nutr.

Trends Pharmacol. Sci.

Selenium in human health and disease

Antioxid. Redox Signal.

In vitro and in vivo studies of methylseleninic acid: evidence that a monomethylated selenium metabolite is critical for cancer chemoprevention

Cancer Res.

Superior in vivo inhibitory efficacy of methylseleninic acid against human prostate cancer over selenomethionine or selenite

Carcinogenesis