Identification of suitable mRNAs and microRNAs as reference genes for expression analyses in skin cells under sex hormone exposure.

Cell Line Gene Expression Profiling Gonadal Steroid Hormones / pharmacology Humans MicroRNAs / genetics RNA, Messenger / genetics Skin / drug effects
Fibroblasts Keratinocytes Melanoma cells Reference genes Sex hormones miRNA

Journal

Gene
ISSN: 1879-0038
Titre abrégé: Gene
Pays: Netherlands
ID NLM: 7706761

Informations de publication

Date de publication:
15 Feb 2021
Historique:
received: 07 05 2020
revised: 02 10 2020
accepted: 27 11 2020
pubmed: 11 12 2020
medline: 30 1 2021
entrez: 10 12 2020
Statut: ppublish

Résumé

Quantitative RT-PCR is the most accurate technique for the study of gene expression profiles, however, to ensure the accuracy of qPCR results, suitable reference genes are necessary for data normalization. Hormones influence the development and function of skin cells, regulating the expression of genes and miRNAs. Nevertheless, the stability of reference genes after sex hormone treatment has not been thoroughly investigated. In this study, we evaluated the expression of a set of candidate mRNAs and microRNsA (miRNA) as reference genes in keratinocytes (HaCaT cells), primary human fibroblasts and a melanoma cell line (LM-36 cells) under testosterone or 17β-estradiol treatment. Two algorithms, namely geNorm, Best-Keeper, and the comparative ΔCt method were used to evaluate the expression stability of the candidate reference genes. The comprehensive ranking showed that TBP and miR-191-5p are the most stable expressed genes across all cultured cells under hormone treatment. Furthermore, we observed that GAPDH, HPRT1 and U6 snRNA expression may be altered by hormone exposure, thus, these genes are not recommended as reference genes. In conclusion, the present study provides, to the best of our knowledge, the first evaluation of expressed mRNA(s) and miRNA(s) as reference genes in three different types of skin cells under the stimulation of sex hormones.

Identifiants

DOI: 10.1016/j.gene.2020.145336 PMID: 33301797
pubmed: 33301797
pii: S0378-1119(20)31005-2
doi: 10.1016/j.gene.2020.145336
pii:
doi:

Substances chimiques

Gonadal Steroid Hormones 0
MicroRNAs 0
RNA, Messenger 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

145336

Informations de copyright

Copyright © 2020 Elsevier B.V. All rights reserved.

Auteurs

S Fochi (S)

Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Italy. Electronic address: stefania.fochi@univr.it.

E Orlandi (E)

Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Italy. Electronic address: elisa.orlandi@univr.it.

L Ceccuzzi (L)

Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Italy. Electronic address: laura.ceccuzzi@univr.it.

M Rodolfo (M)

Melanoma and Sarcoma Surgery Unit and Unit of Immunotherapy, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy. Electronic address: Monica.Rodolfo@istitutotumori.mi.it.

E Vergani (E)

Melanoma and Sarcoma Surgery Unit and Unit of Immunotherapy, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy. Electronic address: elisabetta.vergani@istitutotumori.mi.it.

A Turco (A)

Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Italy. Electronic address: alberto.turco@univr.it.

M G Romanelli (MG)

Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Italy. Electronic address: mariagrazia.romanelli@univr.it.

M Gomez-Lira (M)

Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Italy. Electronic address: macarena.gomezlira@univr.it.

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Classifications MeSH

questionsmedicales.fr › Cellules › Cellules cultivées › Lignée cellulaire › Identification of suitable mRNAs and microRNAs...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Analyse de profil d'expression de gènes › Identification of suitable mRNAs and microRNAs...
questionsmedicales.fr › Hormones gonadiques › Hormones sexuelles stéroïdiennes › Identification of suitable mRNAs and microRNAs...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Identification of suitable mRNAs and microRNAs...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN non traduit › Petit ARN non traduit › microARN › Identification of suitable mRNAs and microRNAs...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN messager › Identification of suitable mRNAs and microRNAs...
questionsmedicales.fr › Système tégumentaire › Peau › Identification of suitable mRNAs and microRNAs...