PMID- 31569378
OWN - NLM
STAT- MEDLINE
DCOM- 20200221
LR  - 20200221
IS  - 2073-4425 (Electronic)
IS  - 2073-4425 (Linking)
VI  - 10
IP  - 10
DP  - 2019 Sep 28
TI  - The Selection and Validation of Reference Genes for mRNA and microRNA Expression 
      Studies in Human Liver Slices Using RT-qPCR.
LID - 10.3390/genes10100763 [doi]
LID - 763
AB  - The selection of a suitable combination of reference genes (RGs) for data 
      normalization is a crucial step for obtaining reliable and reproducible results 
      from transcriptional response analysis using a reverse transcription-quantitative 
      polymerase chain reaction. This is especially so if a three-dimensional 
      multicellular model prepared from liver tissues originating from biologically 
      diverse human individuals is used. The mRNA and miRNA RGs stability were studied 
      in thirty-five human liver tissue samples and twelve precision-cut human liver 
      slices (PCLS) treated for 24 h with dimethyl sulfoxide (controls) and PCLS 
      treated with beta-naphthoflavone (10 microM) or rifampicin (10 microM) as cytochrome P450 
      (CYP) inducers. Validation of RGs was performed by an expression analysis of 
      CYP3A4 and CYP1A2 on rifampicin and beta-naphthoflavone induction, respectively. 
      Regarding mRNA, the best combination of RGs for the controls was YWHAZ and B2M, 
      while YWHAZ and ACTB were selected for the liver samples and treated PCLS. 
      Stability of all candidate miRNA RGs was comparable or better than that of 
      generally used short non-coding RNA U6. The best combination for the control PCLS 
      was miR-16-5p and miR-152-3p, in contrast to the miR-16-5b and miR-23b-3p 
      selected for the treated PCLS. Our results showed that the candidate RGs were 
      rather stable, especially for miRNA in human PCLS.
FAU - Zarybnicky, Tomas
AU  - Zarybnicky T
AUID- ORCID: 0000-0001-6402-9708
AD  - Department of Biochemical Sciences, Charles University, Faculty of Pharmacy in 
      Hradec Kralove, 500 05 Hradec Kralove, Czech Republic. zarybnto@faf.cuni.cz.
FAU - Matouskova, Petra
AU  - Matouskova P
AUID- ORCID: 0000-0002-9421-5744
AD  - Department of Biochemical Sciences, Charles University, Faculty of Pharmacy in 
      Hradec Kralove, 500 05 Hradec Kralove, Czech Republic. matousp7@faf.cuni.cz.
FAU - Ambroz, Martin
AU  - Ambroz M
AD  - Department of Biochemical Sciences, Charles University, Faculty of Pharmacy in 
      Hradec Kralove, 500 05 Hradec Kralove, Czech Republic. ambrozm@faf.cuni.cz.
FAU - Subrt, Zdenek
AU  - Subrt Z
AD  - Department of General Surgery, Third Faculty of Medicine and University Hospital 
      Kralovske Vinohrady, Charles University, 100 34 Prague, Czech Republic. 
      subrt@email.cz.
AD  - Department of Surgery, University Hospital Hradec Kralove, 500 05 Hradec Kralove, 
      Czech Republic. subrt@email.cz.
FAU - Skalova, Lenka
AU  - Skalova L
AD  - Department of Biochemical Sciences, Charles University, Faculty of Pharmacy in 
      Hradec Kralove, 500 05 Hradec Kralove, Czech Republic. skaloval@faf.cuni.cz.
FAU - Bousova, Iva
AU  - Bousova I
AUID- ORCID: 0000-0003-2863-717X
AD  - Department of Biochemical Sciences, Charles University, Faculty of Pharmacy in 
      Hradec Kralove, 500 05 Hradec Kralove, Czech Republic. Iva.Bousova@faf.cuni.cz.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190928
PL  - Switzerland
TA  - Genes (Basel)
JT  - Genes
JID - 101551097
RN  - 0 (14-3-3 Proteins)
RN  - 0 (B2M protein, human)
RN  - 0 (MicroRNAs)
RN  - 0 (RNA, Messenger)
RN  - 0 (YWHAZ protein, human)
RN  - 0 (beta 2-Microglobulin)
RN  - 6051-87-2 (beta-Naphthoflavone)
RN  - 9035-51-2 (Cytochrome P-450 Enzyme System)
RN  - EC 1.14.14.1 (CYP1A2 protein, human)
RN  - EC 1.14.14.1 (Cytochrome P-450 CYP1A2)
RN  - EC 1.14.14.1 (Cytochrome P-450 CYP3A)
RN  - EC 1.14.14.55 (CYP3A4 protein, human)
RN  - VJT6J7R4TR (Rifampin)
RN  - YOW8V9698H (Dimethyl Sulfoxide)
SB  - IM
MH  - 14-3-3 Proteins/genetics/metabolism
MH  - Adult
MH  - Aged
MH  - Cytochrome P-450 CYP1A2/genetics/metabolism
MH  - Cytochrome P-450 CYP3A/genetics/metabolism
MH  - Cytochrome P-450 Enzyme System/pharmacology
MH  - Dimethyl Sulfoxide/pharmacology
MH  - Female
MH  - Gene Expression Profiling/*standards
MH  - Humans
MH  - Liver/drug effects/*metabolism
MH  - Male
MH  - MicroRNAs/*genetics/metabolism
MH  - Middle Aged
MH  - RNA, Messenger/*genetics/metabolism
MH  - Real-Time Polymerase Chain Reaction/*standards
MH  - Reference Standards
MH  - Rifampin/pharmacology
MH  - Transcriptome
MH  - beta 2-Microglobulin/genetics/metabolism
MH  - beta-Naphthoflavone/pharmacology
PMC - PMC6826422
OTO - NOTNLM
OT  - RT-qPCR
OT  - human liver
OT  - mRNA
OT  - miRNA
OT  - precision-cut liver slices
OT  - reference gene
COIS- The authors declare no conflicts of interest.
EDAT- 2019/10/02 06:00
MHDA- 2020/02/23 06:00
PMCR- 2019/10/01
CRDT- 2019/10/02 06:00
PHST- 2019/08/29 00:00 [received]
PHST- 2019/09/25 00:00 [revised]
PHST- 2019/09/27 00:00 [accepted]
PHST- 2019/10/02 06:00 [entrez]
PHST- 2019/10/02 06:00 [pubmed]
PHST- 2020/02/23 06:00 [medline]
PHST- 2019/10/01 00:00 [pmc-release]
AID - genes10100763 [pii]
AID - genes-10-00763 [pii]
AID - 10.3390/genes10100763 [doi]
PST - epublish
SO  - Genes (Basel). 2019 Sep 28;10(10):763. doi: 10.3390/genes10100763.