ShRNA-mediated gene silencing of AHR promotes the differentiation of P19 mouse embryonic carcinoma cells into cardiomyocytes

Mol Med Rep. 2012 Sep;6(3):513-8. doi: 10.3892/mmr.2012.941. Epub 2012 Jun 8.

Abstract

The aryl hydrocarbon receptor (AHR) is a basic helix-loop-helix (bHLH) transcription factor that is activated by environmental contaminants including polychlorinated biphenyls (PCBs). The AHR affects a variety of processes that are involved in cell growth and differentiation. In this study, we constructed a P19 embryonic carcinoma cell line with AHR gene silencing using the vector-based approach of short hairpin (sh)RNA interference that allows cells to differentiate into cardiac myocytes when treated with dimethyl sulfoxide (DMSO). The expression levels of the cardiac development-specific GATA4 and Nkx2.5 genes were measured using real-time quantitative polymerase chain reaction (qPCR). Our data showed that the expression levels of the GATA4 and Nkx2.5 genes were increased in the AHR-silenced P19 cells compared with the control groups. Four critical genes (ARNT, CYP1A1, GSK3β and β-catenin) expressed in the AHR and in the Wnt signaling pathway were also measured by qPCR. We found that the expression levels of ARNT, CYP1A1 and β-catenin were suppressed, whereas GSK3β expression was elevated in the AHR-silenced P19 cells. Therefore, it is possible that the silencing of AHR promotes the differentiation of P19 cells through the AHR and Wnt signal transduction pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aryl Hydrocarbon Receptor Nuclear Translocator / genetics
  • Aryl Hydrocarbon Receptor Nuclear Translocator / metabolism
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Cytochrome P-450 CYP1A1 / genetics
  • Cytochrome P-450 CYP1A1 / metabolism
  • Dimethyl Sulfoxide / pharmacology
  • Embryonal Carcinoma Stem Cells / cytology*
  • Embryonal Carcinoma Stem Cells / metabolism
  • GATA4 Transcription Factor / genetics
  • GATA4 Transcription Factor / metabolism
  • Glycogen Synthase Kinase 3 / genetics
  • Glycogen Synthase Kinase 3 / metabolism
  • Glycogen Synthase Kinase 3 beta
  • Homeobox Protein Nkx-2.5
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Mice
  • Myocytes, Cardiac / cytology*
  • RNA Interference*
  • RNA, Small Interfering / metabolism*
  • Receptors, Aryl Hydrocarbon / antagonists & inhibitors
  • Receptors, Aryl Hydrocarbon / genetics
  • Receptors, Aryl Hydrocarbon / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Wnt Signaling Pathway / drug effects
  • beta Catenin / genetics
  • beta Catenin / metabolism

Substances

  • Arnt protein, mouse
  • GATA4 Transcription Factor
  • Gata4 protein, mouse
  • Homeobox Protein Nkx-2.5
  • Homeodomain Proteins
  • Nkx2-5 protein, mouse
  • RNA, Small Interfering
  • Receptors, Aryl Hydrocarbon
  • Transcription Factors
  • beta Catenin
  • Aryl Hydrocarbon Receptor Nuclear Translocator
  • Cytochrome P-450 CYP1A1
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • Glycogen Synthase Kinase 3
  • Dimethyl Sulfoxide