Elsevier

Journal of Functional Foods

Volume 34, July 2017, Pages 36-48
Journal of Functional Foods

Antiproliferative effect of peptide fractions isolated from a quality protein maize, a white hybrid maize, and their derived peptides on hepatocarcinoma human HepG2 cells

https://doi.org/10.1016/j.jff.2017.04.015Get rights and content

Highlights

  • Albumins in varieties of maize provide reservoir of bioactive peptides.

  • Peptides from albumin maize inhibit cell growth by apoptosis of HepG2 cells.

  • Maize peptides decrease antiapoptotic factors in HepG2 cells.

  • Peptide sequences have potential application as ingredients or supplements.

Abstract

Hydrolysates and peptides isolated from food proteins have shown antiproliferative effects on cancer cells. Cereals are one of the main sources of protein in the diet; among cereals, maize is the most extensively cultivated and consumed in the world. The aim was to compare the antiproliferative activity of ionic exchange peptide fractions isolated from albumin alcalase hydrolysates of white hybrid (Asgrow-773) and quality protein (CML-502) maize, and their derived peptides, using an in vitro model of human liver cancer. Fraction 10 from Asgrow-773 (IC50 = 8.9 µg/mL) was more potent (p < 0.05) than the one from CML-502 (IC50 = 15.7 µg/mL). The treatment with fraction 10 from both genotypes increased apoptosis induction rates an average of 4-fold in HepG2 cells. These results suggest that the antiproliferative effect on HepG2 cells of peptide fractions isolated from both genotypes, but not pure peptides, was based on induction of apoptosis due to decrease of antiapoptotic factors expression.

Introduction

Worldwide, primary liver cancer is the sixth most common cancer and the second cause of cancer-related death (Ferlay et al., 2015). The main risk factors for the development of liver cancer are chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV), obesity, type 2 diabetes, alcohol consumption, cirrhosis, non-alcoholic fatty liver disease, smoking and exposure to aflatoxins (Bisteau et al., 2014, Torre et al., 2016, World Cancer Research Fund & American Institute for Cancer Research, 2007). The number of cancer cases and deaths is expected to grow rapidly as populations grow, age, and adopt lifestyle behaviours that increase cancer risk (Torre et al., 2016).

Several epidemiological studies have shown a correlation between the consumption of whole grains, and their products, with the reduced incidence of chronic diseases, such as cardiovascular disease, type II diabetes and some types of cancer (Ortiz-Martinez, Winkler, & García-Lara, 2014). Cereals are one of the main sources of protein in the diet of people in developing countries because of their availability and low price, compared with other protein-rich sources (Ignjatovic-Micic et al., 2015, Nuss and Tanumihardjo, 2010, Pechanova et al., 2013).

Food proteins are a relevant source of bioactive peptides encrypted in their protein sequence, released by enzymatic hydrolysis or gastrointestinal digestion (Cavazos and Gonzalez de Mejia, 2013, Eckert et al., 2013). Within cereals, one of the most extensively cultivated and consumed in the world is maize (Zea mays L.) (FAO, 2016).

Quality Protein Maize (QPM) genotypes were first developed in a biofortification program lead by S.K. Vasil and E. Villegas at CIMMYT (International Maize and Wheat Improvement Center) in Mexico (Shewry, 2007). QPM genotypes of maize are the product of the opaque-2/ floury mutation coupled with modifications for the restoration of vitreous endosperm (Rosales et al., 2011, Shewry, 2007). QPM genotypes have a higher protein quantity and protein quality than other currently available commercial hybrids. This difference is due to a different distribution of its proteins, with an increase in albumins and globulins, a decrease in prolamins, and in some cases, a higher content of glutelins, as well as marked differences in their amino acid composition (Nuss and Tanumihardjo, 2011, Wu et al., 2010). Beyond the protein biofortification, QPM maize has shown nutritional beneficial effects because it may influence the incorporation of minerals and cofactors into the diet (Nuss and Tanumihardjo, 2011, Rosales et al., 2011).

Bioactive peptides isolated from corn and their industrial co-products have shown to exert a wide variety of biological effects acting as antioxidants (Jin et al., 2016, Wang et al., 2015, Zhou et al., 2015), enzyme inhibitors (Huang, Sun, He, Dong, & Li, 2011), chemo protectors (Lv et al., 2013, Ma et al., 2015, Wu et al., 2014), growth inhibitors and inducers of apoptosis on cancer cells (Li et al., 2013). However, there is a need of knowledge on the differential protein profile of QPM maize and the bioactivity of its proteins, hydrolysates, and isolated peptides. The hypothesis of this research was that regardless of their differential protein and amino acid composition, both the non-protein enriched maize and the QPM, but not pure peptides, may exert antiproliferative effects in human liver cancer cells (HepG2) by induction of apoptosis and arresting of the cell cycle. Therefore, the aim of this research was to compare the antiproliferative activity of ionic exchange peptide fractions isolated from albumin alcalase hydrolysates of white hybrid maize (Asgrow-773) and QPM (CML-502) maize, and their derived peptides, using an in vitro model of human liver cancer.

Section snippets

Materials and reagents

Asgrow-773 (non-protein-enriched, white dent maize) and CML-502 (second-generation QPM inbred line) maize genotypes (Shewry, 2007) were provided by INIFAP (Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Mexico). Ethanol was purchased from VWR BDH-6014-4 (VWR International, USA). Syringe filters were from Pall Corporation. XK 16/20 column and Q-Sepharose Fast Flow resin for the ionic exchange were purchased from GE Healthcare Life Sciences (Buckinghamshire, UK). The

Peptide sequence identification of the IEX maize fractions

The sequences, physicochemical properties and biological potential of the main peptides present in the fraction 10 from Asgrow-773 and CML-502 maize are listed in Table 1, Table 2, respectively. Hydrophobicity of the peptides from Asgrow-773 ranged from +8.43 to 15.53 kcal/mol−1; for the peptides from the CML-502 the range was wider from +8.17 to 20.77 kcal/mol−1. It was noticeable that there were not the same peptide sequences present in both genotypes. It was expected that the resulting

Conclusions

To our understanding, this is the first report of the antiproliferative activity of alcalase hydrolysates of albumin from selective maize genotypes on liver cancer cells. The results obtained in this work demonstrated that fractions isolated from albumins of maize have an antiproliferative effect on HepG2 cells. Fraction 10 isolated from both genotypes were capable of inducing apoptosis, affecting the pattern of expression of anti-apoptotic factors such as livin, survivin, cIAP, XIAP, and

Conflict of interest

The authors declare no conflict of interest.

Acknowledgements

Authors thank NutriOmics from Tecnológico de Monterrey and University of Illinois for the financial support, Consejo Nacional de Ciencia y Tecnologia, CONACyT-Mexico for the scholarship received by Margarita Ortiz-Martinez (number 346902) and Dr. Ricardo Ernesto Preciado Ortiz from INIFAP for providing the maize samples for this study.

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