DOI: 10.1101/469304Nov 14, 2018Paper

Exposure to graphene oxide sheets alters the expression of reference genes used for real-time RT-qPCR normalization

BioRxiv : the Preprint Server for Biology
Irene de Lazaro, Kostas Kostarelos

Abstract

Background: studies that unravel the interactions between thin, 2D graphene oxide (GO) sheets and the biological milieu, including cells and tissues, are multiplying quickly as the biomedical applications of those and other 2D materials continue to be explored. Many of such studies rely on real-time RT-qPCR as a powerful, yet relatively simple technique to determine gene expression. However, a systematic investigation of potential GO-induced changes in the expression of reference genes, crucial for appropriate normalization of qPCR data that ensures reliability of the results, is still lacking. In this study, we aimed to cover this gap by investigating the stability of the expression of ten (10) candidate reference genes upon exposure to increasing, but subtoxic, concentrations of GO, with two established algorithms (Bestkeeper and NormFinder). The study was performed in a human cancer cell line (MCF7) and in mouse, non-cancerous primary cells (mouse embryonic fibroblasts, MEFs), to assess different behaviors between cell types. Results: Bestkeeper and NormFinder algorithms evidenced significant deviations in the expression of various reference genes. Ribosomal proteins scored among the most significantly dysregulated targets i...Continue Reading

Related Concepts

Gene Expression
Genes
Laboratory mice
Ribosomal Proteins
Cell Line, Tumor
Surgical Drapes
Tumor Cells, Malignant
MCF-7 Cells
Protein Expression
Elf4 protein, mouse

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