Antiproliferative Effect of Elastin-Derived Peptide VGVAPG on SH-SY5Y Neuroblastoma Cells

Konrad A Szychowski,Agnieszka Dołhańczuk-Śródka,Agnieszka Rombel-Bryzek,Jan Gmiński

doi:10.1007/s12640-019-00040-y

Konrad A Szychowski, Agnieszka Dołhańczuk-Śródka + Show 2 more

Open Access

https://doi.org/10.1007/s12640-019-00040-y

Copy DOI

Journal: Neurotoxicity Research	Publication Date: Jun 3, 2019
Citations: 17	License type: open-access

Affiliation: Opole University

Abstract

Throughout the lifetime of humans, the amount of stem cells and the rate of cell proliferation continue to decrease. Reactive oxygen species (ROS) are one among the many factors that promote stem cell aging. Both a decrease in the level of stem cells and increase in ROS production can lead to the development of different neurodegenerative diseases. This study was conducted to determine how the VGVAPG peptide, liberated from elastin during the aging process and under pathological conditions, affects ROS production and activities of antioxidant enzymes in undifferentiated, proliferating SH-SY5Y cells. SH-SY5Y cells were maintained in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 supplemented with 10% heat-inactivated fetal bovine serum (FBS). After treating the SH-SY5Y cells with VGVAPG peptide, we measured ROS production; cell metabolism, proliferation, and expression; and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). We demonstrated that the VGVAPG peptide increases GPx expression and activity, whereas it decreases CAT expression in SH-SY5Y cells. Silencing of the GLB1 gene prevents changes in GPx activity. Despite the fact that the VGVAPG peptide increases GPx expression, it increases the ROS level. Moreover, the VGVAPG peptide decreases SH-SY5Y proliferation, which is prevented by the ROS scavenger N-acetyl-L-cysteine. Our data suggest that ROS production and decreased proliferation of SH-SY5Y cells are the results of excitotoxicity meditated through close unrecognized molecular pathways. More research is needed to elucidate the unknown mechanism of action of VGVAPG peptide in the nervous system.

Highlights

Throughout the human lifetime, the amount of stem cells and rate of cell proliferation continue to decrease (Apple et al 2017)
After the SH-SY5Y cells were exposed to concentration of 100 pM to 100 μM VGVAPG for 3 h only in the range 50 nM to 50 μM, respectively, samples with the VGVAPG concentration showed increased reactive oxygen species (ROS) production by 33.98 to 37.82% compared to the control (Fig. 1a)
ROS production increased by 44.45 to 47.25% compared to the control (Fig. 1b) after SH-SY5Y cells were exposed for 6 h to concentrations of 10 nM to 50 μM VGVAPG

Summary

Introduction

Throughout the human lifetime, the amount of stem cells and rate of cell proliferation continue to decrease (Apple et al 2017). It has been described that the decrease in stem cell level and increase in ROS production can both lead to the development of different neurodegenerative diseases (Kim et al 2015). ROS damages the cells and walls of brain capillaries, and this leads to the release of elastin-derived peptides (EDPs) from the extracellular matrix (ECM) (Nita and Grzybowski 2016). It is well documented that the VGVAPG sequence (alone or as a part of EDPs) binds with high affinity to elastin-binding protein (EBP) located on the cell surface (Senior et al 1984; Blood et al 1988). The second receptor for the VGVAPG peptide is galectin-3, which has an important role in cell–ECM interactions (Ochieng et al 2002). Galectin is mostly expressed in inflammatory cells (Bresalier et al 1996; Cantarelli et al 2009); its expression has been linked with tumor progression, cancer aggressiveness, and melanoma invasiveness (Ochieng et al 1999; Pocza et al 2008; Wang et al 2009)

Methods

Results

Discussion

Conclusion