Document Detail


Normalization genes for quantitative RT-PCR in differentiated Caco-2 cells used for food exposure studies.
MedLine Citation:
PMID:  21779557     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Exposure of food products to small-intestinal-like Caco-2 cells, combined with a gene expression based response analysis can be a valuable tool to classify potential bioactive effects of food homogenates. In order to study changes in gene expression upon food exposure, a robust set of stably expressed genes is required for normalization. Here we present a set of reference genes suitable for RT-qPCR that has been validated for exposure studies with the intestinal-like Caco-2 cell line. This study identified ribosomal phosphoprotein P0 (RPLP0) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as best reference genes. The set can be extended with β-2-microglobulin (B2M), splicing factor 3A, subunit 1 (SF3A1), and mitochondrial ribosomal protein L19 (MRPL19). Food homogenates did provoke responses in the Caco-2 cells, as was demonstrated by changed expression of NAD(P)H Quinone dehydrogenase 1 (NQO1), Claudin 4 (CLDN4), Nitric Oxide Synthase 2 (NOS2), and ATP-binding cassette, subfamily B, member 1 (ABCB1) in the same experiment. Results indicate that: i) natural food homogenates can exert effects in Caco-2 cells, and ii) stability in expression of the reference genes is not due to a lack of response of the Caco-2 cells.
Authors:
Robert A M Vreeburg; Shanna Bastiaan-Net; Jurriaan J Mes
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Validation Studies     Date:  2010-12-13
Journal Detail:
Title:  Food & function     Volume:  2     ISSN:  2042-650X     ISO Abbreviation:  Food Funct     Publication Date:  2011 Feb 
Date Detail:
Created Date:  2011-07-22     Completed Date:  2012-01-16     Revised Date:  2012-05-28    
Medline Journal Info:
Nlm Unique ID:  101549033     Medline TA:  Food Funct     Country:  England    
Other Details:
Languages:  eng     Pagination:  124-9     Citation Subset:  IM    
Affiliation:
Wageningen University and Research centre, Food & Biobased Research, P.O. Box 17, 6700 AA, Wageningen, The Netherlands. robert.vreeburg@wur.nl
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MeSH Terms
Descriptor/Qualifier:
Caco-2 Cells
Claudins / genetics
DNA Primers / standards*
Food*
Fruit*
Gene Expression / genetics
Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
Humans
Mitochondrial Proteins / genetics
NAD(P)H Dehydrogenase (Quinone) / genetics
Nitric Oxide Synthase Type II / genetics
P-Glycoprotein / genetics
Real-Time Polymerase Chain Reaction / methods,  standards*
Reference Standards
Ribonucleoprotein, U2 Small Nuclear / genetics
Ribosomal Proteins / genetics
Vegetables*
beta 2-Microglobulin / genetics
Chemical
Reg. No./Substance:
0/ABCB1 protein, human; 0/Claudins; 0/DNA Primers; 0/MRPL19 protein, human; 0/Mitochondrial Proteins; 0/P-Glycoprotein; 0/Ribonucleoprotein, U2 Small Nuclear; 0/Ribosomal Proteins; 0/SF3A1 protein, human; 0/beta 2-Microglobulin; 0/claudin 4; 0/ribosomal protein P0; EC 1.14.13.39/NOS2 protein, human; EC 1.14.13.39/Nitric Oxide Synthase Type II; EC 1.2.1.-/Glyceraldehyde-3-Phosphate Dehydrogenases; EC 1.6.5.2/NAD(P)H Dehydrogenase (Quinone); EC 1.6.5.2/NQO1 protein, human

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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