PMID- 21779557
OWN - NLM
STAT- MEDLINE
DCOM- 20120116
LR  - 20191210
IS  - 2042-650X (Electronic)
IS  - 2042-6496 (Linking)
VI  - 2
IP  - 2
DP  - 2011 Feb
TI  - Normalization genes for quantitative RT-PCR in differentiated Caco-2 cells used 
      for food exposure studies.
PG  - 124-9
LID - 10.1039/c0fo00068j [doi]
AB  - 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 beta-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.
FAU - Vreeburg, Robert A M
AU  - Vreeburg RA
AD  - Wageningen University and Research centre, Food & Biobased Research, P.O. Box 17, 
      6700 AA, Wageningen, The Netherlands. robert.vreeburg@wur.nl
FAU - Bastiaan-Net, Shanna
AU  - Bastiaan-Net S
FAU - Mes, Jurriaan J
AU  - Mes JJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20101213
PL  - England
TA  - Food Funct
JT  - Food & function
JID - 101549033
RN  - 0 (ABCB1 protein, human)
RN  - 0 (ATP Binding Cassette Transporter, Subfamily B)
RN  - 0 (ATP Binding Cassette Transporter, Subfamily B, Member 1)
RN  - 0 (CLDN4 protein, human)
RN  - 0 (Claudin-4)
RN  - 0 (Claudins)
RN  - 0 (DNA Primers)
RN  - 0 (MRPL19 protein, human)
RN  - 0 (Mitochondrial Proteins)
RN  - 0 (RNA Splicing Factors)
RN  - 0 (Ribonucleoprotein, U2 Small Nuclear)
RN  - 0 (Ribosomal Proteins)
RN  - 0 (SF3A1 protein, human)
RN  - 0 (beta 2-Microglobulin)
RN  - 0 (ribosomal protein P0)
RN  - EC 1.14.13.39 (NOS2 protein, human)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase Type II)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
RN  - EC 1.6.5.2 (NAD(P)H Dehydrogenase (Quinone))
RN  - EC 1.6.5.2 (NQO1 protein, human)
SB  - IM
MH  - ATP Binding Cassette Transporter, Subfamily B
MH  - ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics
MH  - Caco-2 Cells
MH  - Claudin-4
MH  - Claudins/genetics
MH  - DNA Primers/*standards
MH  - *Food
MH  - *Fruit
MH  - Gene Expression/genetics
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/genetics
MH  - Humans
MH  - Mitochondrial Proteins/genetics
MH  - NAD(P)H Dehydrogenase (Quinone)/genetics
MH  - Nitric Oxide Synthase Type II/genetics
MH  - RNA Splicing Factors
MH  - Real-Time Polymerase Chain Reaction/methods/*standards
MH  - Reference Standards
MH  - Ribonucleoprotein, U2 Small Nuclear/genetics
MH  - Ribosomal Proteins/genetics
MH  - *Vegetables
MH  - beta 2-Microglobulin/genetics
EDAT- 2011/07/23 06:00
MHDA- 2012/01/17 06:00
CRDT- 2011/07/23 06:00
PHST- 2011/07/23 06:00 [entrez]
PHST- 2011/07/23 06:00 [pubmed]
PHST- 2012/01/17 06:00 [medline]
AID - 10.1039/c0fo00068j [doi]
PST - ppublish
SO  - Food Funct. 2011 Feb;2(2):124-9. doi: 10.1039/c0fo00068j. Epub 2010 Dec 13.