Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCA267

Poster Communications

The effect of rhein on protein expression and differentiation of SH-SY5Y neuroblastoma cells

Z. Cockova1, H. Ujcikova1,2, P. Telensky1,3, J. Novotny1

1. Department of Physiology, Faculty of Science, Charles University, Prague, Czechia. 2. Institute of Physiology, Czech Academy of Sciences, Prague, Czechia. 3. International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czechia.

Apart from its connection to the tumorous character, disrupted energy metabolism may be linked to the progression of neurodegenerative diseases. Rhein, a potential candidate for preventing metabolic disorders, has been shown to have various pharmacological properties including anticancer activity and modulatory effects on the bioenergetics. However, its effects on neuronal metabolism are poorly understood. We sought to analyze the impact of rhein treatment on protein expression and the differentiation process of SH-SY5Y neuroblastoma cells. Using two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry we evaluated changes in the proteome of both undifferentiated and retinoic acid-differentiated SH-SY5Y cells after 24-h treatment with rhein. This analysis led to identification of 7 differentially expressed proteins in samples from undifferentiated cells (actin, cytoplasmic 1; tubulin beta chain; tubulin alpha-1A chain; alpha-centractin; protein disulfide-isomerase A3; heterogeneous nuclear ribonucleoprotein A1; 3-ketoacyl-CoA thiolase) and 10 differentially expressed proteins in samples from differentiated SH-SY5Y cells (actin, cytoplasmic 1; tubulin beta chain; tubulin alpha-1B chain; stress-70 protein; heterogeneous nuclear ribonucleoprotein H; thioredoxin-dependent peroxide reductase; peroxiredoxin-6; heterogeneous nuclear ribonucleoprotein L; poly(rC)-binding protein 1; fructose-bisphosphate aldolase A). All of them were downregulated at least 2-fold (p<0.05; Student's t-test) by treatment with rhein. These proteins are involved in cellular metabolism, cytoskeleton organization, transcriptional and translational regulation and antioxidant defence. Validation of selected differentially expressed proteins performed by Western blotting revelaed identical trends in the expression of these proteins (n=5). Addition of rhein to the cultured cells during differentiation resulted in an altered expression of the following 3 markers: microtubule-associated protein 2, nestin and growth associated protein 43. Relative levels of selected proteins were determined by Western blotting (n=5). In parallel, rhein significantly reduced neurite outgrowth (136.8 ± 4.0 µm vs 205.3 ± 6.7 µm; p<0.001; Student's t-test). Values are expressed as means ± S.E.M. Neurite outgrowth was evaluated by tracing neurites (n=70) on microscopy images using ImageJ NeuronGrowth plugin. Our results indicate that rhein may strongly interfere with the differentiation process of SH-SY5Y neuroblastoma cells and is capable of inducing marked proteomic changes in these cells. The results also demonstrate that rhein-treated neuroblastoma cells display dysregulation of proteins involved in cytoskeleton organization, protein folding, metabolism and transcription control.

Where applicable, experiments conform with Society ethical requirements