Universität zu Köln

del Pozo, Talia, Gutierrez-Garcia, Ricardo, Latorre, Mauricio, Gonzalez, Mauricio and Suazo, Miriam (2016). Identification of reference genes for quantitative real-time PCR studies in human cell lines under copper and zinc exposure. Biometals, 29 (5). S. 935 - 945. DORDRECHT: SPRINGER. ISSN 1572-8773

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Abstract

Accurate quantification depends on normalization of the measured gene expression data. In particular, gene expression studies with exposure to metals are challenging due their toxicity and redox-active properties. Here, we assessed the stability of potential reference genes in three cell lines commonly used to study metal cell metabolism: Caco-2 (colon), HepG2 (liver) and THP-1 (peripheral blood) under copper (Cu) or zinc (Zn) exposure. We used combined statistical tools to identify the best reference genes from a set of eleven candidates, which included traditional housekeeping genes such as GAPDH and B-ACTIN, in cell lines exposed to high and low, Zn and Cu concentrations. The expression stabilities of ATP5B (ATP synthase) and CYC1 (subunits of the cytochrome) were the highest considering the effect of Zn and Cu treatments whereas SDHA (succinate dehydrogenase) was found to be the most unstable gene. Even though the transcriptional effect of Zn and Cu is very different in term of redox properties, the same best reference genes were identified when Zn or Cu treatments were analyzed together. Our results indicate that ATP5B/CYC1 are the best candidates for reference genes after metal exposure, which can be used as a suitable starting point to evaluate gene expression with other metals or in different cell types in human models.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code del Pozo, Talia UNSPECIFIED UNSPECIFIED UNSPECIFIED Gutierrez-Garcia, Ricardo UNSPECIFIED UNSPECIFIED UNSPECIFIED Latorre, Mauricio UNSPECIFIED UNSPECIFIED UNSPECIFIED Gonzalez, Mauricio UNSPECIFIED UNSPECIFIED UNSPECIFIED Suazo, Miriam UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-260351
DOI:	10.1007/s10534-016-9965-x
Journal or Publication Title:	Biometals
Volume:	29
Number:	5
Page Range:	S. 935 - 945
Date:	2016
Publisher:	SPRINGER
Place of Publication:	DORDRECHT
ISSN:	1572-8773
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language RT-PCR; HOUSEKEEPING GENES; MONONUCLEAR-CELLS; MICROARRAY DATA; MESSENGER-RNA; EXPRESSION; NORMALIZATION; VALIDATION; ARABIDOPSIS; SELECTION Multiple languages Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/26035