The Genotoxic and Pro-Apoptotic Activities of Advanced Glycation End-Products (MAGE) Measured with Micronuclei Assay Are Inhibited by Their Low Molecular Mass Counterparts.

Monika Czech,Maria Konopacka,Łukasz Łaczmański,Zbigniew Maniakowski,Magdalena Staniszewska,Danuta Witkowska,Andrzej Gamian,Jacek Rogoliński

doi:10.3390/genes12050729

Abstract

An association between the cancer invasive activities of cells and their exposure to advanced glycation end-products (AGEs) was described early in some reports. An incubation of cells with BSA–AGE (bovine serum albumin–AGE), BSA–carboxymethyllysine and BSA–methylglyoxal (BSA–MG) resulted in a significant increase in DNA damage. We examined the genotoxic activity of new products synthesized under nonaqueous conditions. These were high molecular mass MAGEs (HMW–MAGEs) formed from protein and melibiose and low molecular mass MAGEs (LMW–MAGEs) obtained from the melibiose and N-α-acetyllysine and N-α-acetylarginine. We have observed by measuring of micronuclei in human lymphocytes in vitro that the studied HMW–MAGEs expressed the genotoxicity. The number of micronuclei (MN) in lymphocytes reached 40.22 ± 5.34 promille (MN/1000CBL), compared to 28.80 ± 6.50 MN/1000 CBL for the reference BSA–MG, whereas a control value was 20.66 ± 1.39 MN/1000CBL. However, the LMW–MAGE fractions did not induce micronuclei formation in the culture of lymphocytes and partially protected DNA against damage in the cells irradiated with X-ray. Human melanoma and all other studied cells, such as bronchial epithelial cells, lung cancer cells and colorectal cancer cells, are susceptible to the genotoxic effects of HMW–MAGEs. The LMW–MAGEs are not genotoxic, while they inhibit HMW–MAGE genotoxic activity. With regard to apoptosis, it is induced with the HMW–MAGE compounds, in the p53 independent way, whereas the low molecular mass product inhibits the apoptosis induction. Further investigations will potentially indicate beneficial apoptotic effect on cancer cells.

Highlights

Advanced glycation end-product (AGE) formation and accumulation in tissues play an important role in diseases related to diabetes and ageing processes [1,2]
Glycation products synthesized in anhydrous conditions (HTG) showed genotoxic properties, dose dependent, because the number of micronuclei increased twice in cells BEAS-2B (Table 4) and A549 (Table 5), and a 3-fold increase was observed in HCT 116 (-/-) cells (Table 6)
The new high molecular mass MAGE synthesized at anhydrous conditions from bovine serum albumin (BSA) and MB glycation with D-melibiose (BSA–mel-II, MB–mel) demonstrated significant genotoxic activity against human peripheral blood lymphocytes and cell lines

Summary

Introduction

Advanced glycation end-product (AGE) formation and accumulation in tissues play an important role in diseases related to diabetes and ageing processes [1,2]. The first step of glycation is the formation of a reversible Schiff base [3,4] This unstable bond undergoes an Amadori rearrangement, creating a more stable, but still partially reversible, Amadori product. This 1-amino-1-deoxyketose with carbonyl function is considered an early product of glycation, a precursor of further compounds formed in the process of glycation. The complex cascade of dehydration, fragmentation, condensation, oxidation and cyclization reactions occurs These reactions yield a variety of largely undefined advanced Maillard reaction end-products, so-called AGEs. In the presence of oxygen, the Amadori products are degraded to form even more reactive dicarbonyl derivatives; the term glycoxidation is used for these reactions [5]. The MAGE cross-reactive autoantibodies were detected in patients with diabetes [16]

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